PT791006E - Polyvinyl benzoyl glycosides as inhibitors of the smooth muscle cell proliferation - Google Patents

Abstract

This invention relates to the use of polyanionic benzylglycosides as smooth muscle cell proliferation inhibitors and as therapeutic compositions for treating diseases and conditions which are characterized by excessive smooth muscle proliferation, such as restenosis. The compounds of this invention are those of formula I <IMAGE> I <IMAGE> wherein each of R1, R2, R3, and R4 are, independently, H, SO3M, or <IMAGE> and each oligosaccharide group contains 1 to 3 sugar groups; M is lithium, sodium, potassium, or ammonium; n is 1 or 2; X is a halogen, lower alkyl having 1 to 6 carbon atoms, or lower alkoxy having 1 to 6 carbon atoms; Y is carbonyl or sulfonyl; Z is alkyl from 1 to 12 carbon atoms, <IMAGE> <IMAGE> <IMAGE> and X is as defined above; or a pharmaceutically acceptable salt thereof.

Description

DESCRIPTION &quot; POLYNIONIC BENZYLGLYCOSIDES AS INHIBITORS OF PROLIFERATION OF SMOOTH MUSCLE CELLS "

This invention relates to polyanionic benzylglycosides and their use as inhibitors of smooth muscle cell proliferation and as therapeutic compositions for the treatment of diseases and conditions which are characterized by excessive proliferation of smooth muscle, such as restenosis.

Background of the Invention

All forms of vascular reconstruction such as angioplasty and &quot; bypass &quot; venous lesions result in a response to the lesion leading ultimately to proliferation of smooth muscle cells (SMC) and, subsequently, deposition of profuse amounts of extracellular matrix (Cloves, AW, Reidy, AJ Vasc Surg 1991, 13, 885 ). These events are also central processes in the pathogenesis of atherosclerosis (Raines EW; Ross R. Br. Heart J. 1993, 69 (Supplement), S 30) as well as transplantation atherosclerosis (Isik, FF; McDonald, TO; Ferguson, M Yamanaka, E., Gondon, Am. J. Pathol 1992, 141, 1139). In the case of restenosis consequent to angioplasty, clinically relevant solutions to control the proliferation of SMCs by pharmacological intervention have so far been elusive (Herrman, JPR, Hermans, WRM, Vos, J, Serruys PW Drugs 1993, 4, 18 and 249). Any successful approach to inhibit SMC proliferation should not interfere with repair of endothelial cells or the normal function of any other cell type (Weissberg, PL; Grainger, DJ; Shanahan, CM, Metcalfe, JC Cardio-2 -

vascular Res. 1993, 27, 1191). Indeed, an important therapeutic consideration is to promote re-endothelialization of the concurrent affected area with inhibition of smooth muscle cell proliferation (Casscells, W. Circulation 1992, 86, 722; Reidy, A., Lidner, V. In Endothelial Cell Dysiunctions, Simionescu N. and Simionescu M., Ed. Plenum Press, NY, (1992, 31).

Heparin glycosaminoglycans and heparan sulfate are endogenous inhibitors of SMC proliferations, so they are capable of promoting the growth of endothelial cells (Castellot, JJ, Wright, TC, Kamovsky, MJ Seminars in Thrombosis and Hemostasis 1987, 13, 489 ; Wright, TN Arteriosclerosis 1989, 9, 1). However, the maximum clinical benefits of heparin, heparin fragments, chemically modified heparin, low molecular weight heparins and other heparins that mimic anionic polysaccharides may be compromised due to other pharmacological incompatibilities (excessive bleeding due to anticoagulant effects in particular) associated with the heterogeneity of the various preparations (Borman, S. Chemical and Engineering News, 1993, June 28, 27); Schmid, K. M .; Preisack, M .; Voelker, W .; Sujatta, M .; Karsch, V. R. Seminars in Thrombosis and Hemostasis 1993, 19 (Suppl 1), 155; Amann, F. W .; Neuenschwander, C .; Meyer, B .; Seminars in Thrombosis and Hemostasis 1993, 19 (Suppl 1), 160; Radhakrishnamurthy, B .; Sharma, C .; Bhandaru, R.R .; Berenson, G. S .; Stanzani, L .; Mastacchi, R. Atherosclerosis, 1986, 60, 141; Maffrand, J.P .; Hervert, Μ. M .; Bemat, A .; Deffeyn, G .; Delevassee, D .; Savi, P .; Pinot, J. J .; Sampol, J. Seminars in Thrombosis and Hemostasis, 1991, 17 (Suppl 2), 186). Because the anticoagulant effects of many of these agents are independent of the antiproliferative activity of SMCs, it would be expected that polyanionic agents which are more homogeneous in composition and with a more defined molecular structure would exhibit a more desirable profile with fewer side effects associated with the above-mentioned polysaccharides anionics. Background Art WO 92/18546 discloses specific sequences of heparin, obtainable in pure form through synthesis or isolation of heparin fragments which exhibit antiproliferative activity of the SMCs. Beta-cyclodextrin tetradecasulfate has been described as an inhibitor of smooth muscle cell proliferation and as an effective restenosis inhibitor (Weisz, PB; Hermann, HC; Joullie, M., Kumor, K., Levine, EM; Macarak , EJ Weiner, DB Angiogenesis: Key Principie - Science - Technology - Medicine - Steiner, R. Weisz, PB, Langer, R. Eds Birkhauser Verlag, Basel Switzerland, 1992, p 107, Hermann, HC Okada, S, Hozakowska, E., Le Veen, RF, Golden, MA, Tomaszewski, JE; Weisz, PB; Bamathan, ES Arteriosclerosis and Thrombosis 1993, 13, 924; Reilly, CF; Fujita, T. McFall, RC Stabilito , II, Wai-si, E., Johnson, RG Drug Development Research 1993, 29, 137). U.S. Pat. 5,019,562 discloses anionic derivatives of cyclodextrins for the treatment of pathological conditions associated with undesired growth of cells or tissue. WO 93/09790 discloses polyanionic derivatives of anti-proliferating cyclodextrins having at least 2 anionic residues per each carbohydrate residue. EP 312087 A2 and EP 312086 A2 disclose antithrombotic and anticoagulant properties of sulfated bis-aldonic acid amides.

U.S. Pat. Nos. 4,431,636, 4,431,637, 4,431,638 and 4,435,387 disclose polysulfated glycosidic derivatives, thio and oxyaryl as modulators of the complement system.

The compounds of the present invention differ from all of the foregoing because the compounds (a) are benzylglycosides which have no structural similarity to heparin, sulfated cyclodextrins or sulfated lactobionic acid dimers, (b) contain no more than three contiguous sugar residues (trisaccharides) and (c) have a defined structure.

Description of the Invention

This invention describes the composition and utility of sulfated benzylglycosides of formula I

wherein each R 1, R 2, R 3 and R 4 are independently H, SO 3 M or a sugar group of the structure:

and each oligosaccharide group having the structure

contains 1 to 3 sugar groups; M is lithium, sodium, potassium or ammonium; n is 1 or 2; X is hydrogen, lower alkyl of 1 to 6 carbon atoms or lower alkoxy of 1 to 6 carbon atoms; Y is carbonyl or sulfonyl; Z is a group of formula - (CH 2) p -, wherein p is an integer from 1 to 12; XQ-

X

X

2 x NN 3 5s

and X is as defined above; or a pharmaceutically acceptable salt thereof.

It will be understood that each of the various occurrences of R1 in any given molecule of Formula I may be the same or different. Also, each of the various occurrences of R2, R3 or R4 may be the same as or different from other occurrences of R2, R3 or R4, respectively. Likewise, each of the various occurrences of Y in any given molecule and each occurrence of n in any given molecule may be the same or different from other occurrences of Y or n, respectively.

It is also understood that lower alkyl groups may have straight or branched chains and may be, for example, methyl, ethyl, propyl, isopropyl or butyl groups. Likewise, lower alkoxy groups may have straight or branched chains and may be, for example, methoxy, propoxy, isopropoxy or butoxy groups.

A more preferred aspect or embodiment of this invention are the compounds of formula I:

X

n o X j-N_Y_z_Y_

OR2 n

I wherein each R 1, R 2, R 3 and R 4 are independently H, SO 3 M or:

OR2 and each oligosaccharide group of structure

-7- contains 1 or 2 sugar groups; M is lithium, sodium, potassium or ammonium; n is 1 or 2; X is hydrogen, halogen, lower alkyl of 1 to 6 carbon atoms or lower alkoxy of 1 to 6 carbon atoms; Y is carbonyl or sulfonyl; Z is an alkyl having 1 to 12 carbon atoms,

Η H

or a pharmaceutically acceptable salt thereof.

The most preferred compounds of this invention are:

Dodecanedioic acid bis {[2-methyl-5- (hepta-0-sulfato-β-D-cellobiosyloxymethyl) -phenyl] amide} or a pharmaceutically acceptable salt thereof;

N, N'-Bis [5- (hepta-O-sulfato-β-D-cellobiosyloxymethyl) -2-methylphenyl] terephthalamide tetradecodicarbonic salt or a pharmaceutically acceptable salt thereof; -8-

Bis {[5- (hepta-O-sulfato-β-D-cellobiosyloxymethyl) -2-methylphenyl] amide} tetradecodicarbonic salt or a pharmaceutically acceptable salt thereof;

N, N'-Bis [5- (hepta-O-sulfato-β-D-cellobiosyloxymethyl) -2-methylphenyl] isophthalamide tetradecodicarbonate salt or a pharmaceutically acceptable salt thereof;

Decaudic acid bis {[3,5-bis (hepta-O-sulfato-β-D-cellobiosyloxymethyl) phenylamide] or a pharmaceutically acceptable salt thereof;

Bis [(3,5-Bis (hepta-O-sulfato-β-D-cellobiosyloxymethyl) phenyl] amide} salt of biphenyl-4,4'-dicarboxylic acid or a pharmaceutically acceptable salt thereof;

Bis ([3,5-bis (hepta-0-sulfato-β-D-lactosyloxymethyl) phenyl] -amide} salt of biphenyl-4,4'-dicarboxylic acid or a pharmaceutically acceptable salt thereof;

Bis ([3,5- (hepta-O-sulfato-β-D-maltosyloxymethyl) phenyl] amide} salt of biphenyl-4,4'-dicarboxylic acid or a pharmaceutically acceptable salt thereof;

Biphenyl-4,4'-dicarboxylic acid bis {[3,5-bis (tetra-O-sulfato-β-D-glucosyloxymethyl) phenyl] amide hexadecassic salt or a pharmaceutically acceptable salt thereof;

Biphenyl-4,4'-dicarboxylic acid bis {[2-chloro-5- (hepta-0-sulfato-β-D-cellobiosyloxymethyl) phenyl] amide} tetradecodicarbonic salt or a pharmaceutically acceptable salt thereof; -9-

Biphenyl-4,4'-dicarboxylic acid bis {[4-chloro-2- (hepta-0-sulfato-β-D-cellobiosyloxymethyl) phenyl] amide} or a pharmaceutically acceptable salt thereof;

N, N'-bis [3,5-bis- (hepta-O-sulfato-β-D-cellobiosyloxymethyl) -phenyl] succinamide octacosodium salt or a pharmaceutically acceptable salt thereof;

N, N'-Bis [3,5-bis (hepta-O-sulfato-β-D-cellobiosyloxymethyl) -phenyl] terephthalamide octacosodium salt or a pharmaceutically acceptable salt thereof;

Biphenyl-4,4'-disulfonic acid bis {[2-methyl-5- (hepta-0-sulfato-β-D-cellobiosyloxymethyl) -phenyl] amide} or a pharmaceutically acceptable salt thereof;

N, N'-Bis [2-methyl-5- (2,3,4,6-tetra-O-sulfato-β-D-glucopyranosiloxymethyl) phenyl] succinamide octassodium salt or a pharmaceutically acceptable salt thereof;

3,3 '- [N, N'-ureido] -bis {N- [2-methyl-5- (hepta-0-sulfato-β-D-maltosyloxymethyl) phenyl]} benzamide or a pharmaceutically acceptable salt thereof;

3,3 '- (N, N'-ureido) -bis - ({N- [2-chloro-5- (hepta-0-sulfato-PD-cellobiosyloxymethyl) phenyl]} benzamide) salt or a salt thereof pharmaceutically acceptable salt thereof;

N, N'-Bis {3- [2-methyl-5- (hepta-0-sulfato-β-D-maltosyloxymethyl) phenylcarbamoyl] phenyl} isophthalamide tetradecodicarbonate salt or a pharmaceutically acceptable salt thereof.

Process of the Invention

The compounds of the present invention are prepared according to the general sequence of reactions outlined in the Scheme below:

OR2

Thus, a glycosyl bromide 1 is coupled with a benzyl alcohol 2 in the presence of a catalyst such as mercury bromide, mercury cyanide, silver triflate or silver perchlorate in an aprotic solvent such as dichloromethane, ether, toluene or nitromethane a temperatures ranging from -40 ° C to room temperature to give glycoside 3. Reduction of the 3-nitro group of 3 can be effected with a reducing agent such as tin chloride is effected in a polar aprotic solvent such as ethyl acetate , from room temperature to reflux, or by catalytic hydrogenation in the presence of a catalyst such as palladium on carbon gives an anilino compound 4. The coupling of 4 with a bis acid chloride or sulfonyl chloride C1-Z-YC1 may be effected in the presence of a base amine such as triethylamine or diisopropylethylamine in an aprotic solvent such as dichloromethane or tetrahydrofuran. The hydrolysis of any acetate groups present in the sugars with a base such as sodium methoxide in methanol or aqueous sodium hydroxide in methanol at room temperature to the reflux temperature, and the sulfation of some or all of the free hydroxyl groups in the sugars can be supplemented with a reagent such as the sulfur trioxide-trimethylamine complex or the sulfur trioxide-trimethylamine complex or sulfur trioxide-pyridine complex in a polar aprotic solvent such as dimethylformamide or dimethylsulfoxide at temperatures ranging from 0 ° C to 100 ° C gives the target compounds I.

Altematively, the sulfate groups can be introduced into compound 3 (after hydrolysis of any acetate groups). The sulfated compound 3 is then reduced to give the sulfate 4 and linked with Cl-Z-YC1 to give I. The invention is also directed to pharmaceutical compositions, and to their manufacture, comprising sulfated benzylglycosides alone or in combination with excipients pharmacologically acceptable). Such compositions are useful for diseases and conditions which are characterized by excessive proliferation of smooth muscle cells, such as restenosis, often caused by reconstructive vascular cramping and transplantation, for example, balloon angioplasty, surgical vascular graft, &quot; bypass &quot; coronary artery surgery and cardiac transplantation. Other disease states in which there is undesired vascular proliferation include hypertension, asthma, and congestive heart failure. The compounds of the invention are thus useful for treating such diseases and conditions.

The compounds of this invention may be administered systemically, for example by intravenous injection, typically ranging from 0.1 to 10 mg / kg / hr for 5 to 30 days, or by subcutaneous injection at a lower dose, by oral administration with a higher dose higher than the intravenous injection. Localized application of the compounds of this invention may also be accomplished transmembrane, transdermally or other topical routes of administration using appropriate sustained release devices, such as carrier matrix, where applicable. The compositions of the invention may be formulated with conventional excipients, such as fillers, disintegrating agents, binders, lubricants, flavoring agents and the like. These are formulated in a conventional manner. It is to be understood that the compounds of this invention may be administered in any form and in any concentration which is effective for the particular receptor. The dosage form and composition and concentration will be determined on an individual basis by the physician or other trained medical professional treating the recipient.

Effects of cell proliferation A. Source of Cells The ability of the compounds of the present invention to inhibit proliferation of smooth muscle cells and to modulate endothelial cell growth was established using cells isolated from the aorta of commercial origin or, for some species, local. Cell lines used in this study include human and porcine aorta smooth muscle cells and human aortic endothelial cells. Human aortic cell lines were obtained from Clonetics Corporation (San Diego).

The porcine aortas were received from a local slaughterhouse. The material was transported on ice. The aorta was scrupulously cleared of fatty tissue and washed with sterile phosphate buffered saline and 2% antibiotic / antimycotic (Gibco catalog # 600-5240 AG). The tissue was then digested in 10-15 mL of &quot; Enzyme Mixture &quot; containing 165 U / ml type I collagenase; 15 U / mL elastase type III; 2 mg / ml BSA; and 0.375 mg / ml soybean trypsin inhibitor followed by incubation at 37øC under 5% CO 2 for 10-15 min. After this treatment, the adventitious outer surface was easily removed by tweezing. The aorta was then cut longitudinally, opened and the endothelial layer removed by scraping. The middle layer of the cells was washed in enzyme solution and placed in a new 100 mm dish with 10 ml of enzyme solution. The aorta was cut using sharp scissors and digested for 2-3 hours at 37øC in 30 ml of fresh enzyme solution. After digestion, the tissue was homogenized using a fire-tipped Pasteur pipette or an eppendorf pipettor with a sterile 200-1000 mL pipetting tip. The suspension was then centrifuged for 10 m. At 8000 rpm and the reservoir resuspended in 4 - 6 mL of fresh medium and placed in 4-6 100 mL vials with vented capsules. The cells were allowed to grow to confluence and separated using 0.25% trypsin. Cells were evaluated for purity and overall quality using antibodies to SMC actin. B. Effects of Compounds on Cell Proliferation Using 3H Thimidine Incorporation.

Cells were assayed at early passages (usually passage 3-7) under sub-confluence conditions. Cultures were grown in 16-mm (24-well) multi-well culture plates in 199 medium supplemented with 10% fetal bovine serum and 2% antibiotic / antimycotic. At the sub-confluence the cells were placed in a defined serum-free medium (AIM-V; Gibco) for 24-48 h before the start of the experimental protocol.

Although the compounds were found to be more efficacious with longer preincubations, experiments were generally initiated with the addition of the H thymidine and serum / growth factor compound to the serum-deprived synchronized cells and the results are reported in this accordingly. Stimuli caused by growth factor and serum were optimized for each cell type.

Compounds were added to each well at a 50-fold dilution (20 æl / well) and the plates were incubated for 24-36 h at 37øC with 5% CO 2. These series were found to be all soluble in H 2 O and thus the compounds tested were initially diluted in H 2 O and serially diluted in the media. Compounds were routinely assayed at 1, 10 and 100 μΜ. As a control, Sigma H-7005 pig intestinal mucosal heparin (H-7005) was routinely tested in all cell preparations at concentrations of 0.1 to 100 pg / ml.

At the end of the experiment, the plates were placed on ice, washed three times with ice-cold PBS and incubated in ice-cold 10% trichloroacetic acid (TCA) for 30 min. to remove the acid soluble proteins. The solution was transferred to scintillation vials containing 0.4 N HCl (500 μl / vial to neutralize NaOH) and each well was washed twice with water (500 μl) making a total volume of 2 ml / vial. - 15-

Data were obtained, in triplicate, for both control and experimental samples. Control data (100%) were obtained from maximally stimulated cells as a result of stimulation of growth factor or serum. Experimental data were obtained from maximally stimulated growth factor or serum and treated with compound. Data were expressed as percent of control from which the IC 50 could be determined. The compounds of the present invention are effective inhibitors of smooth muscle cell proliferation as summarized in Table I. In addition, the compounds of the present invention exhibit antiproliferative activity of human smooth muscle cells (HAOSMC) in the case where proliferation is motivated by 10% fetal bovine serum (FBS) or platelet derived growth factor (PDGF; human recombinant PDGF-AB purchased from Upstate Biotechnology Inc., Lake Placid, NY). For example, the sulfated compound of example 18 inhibits the proliferation of FBS-induced HAOSMC (200 nM IC50) as well as 5 ng / ml PDGF (380 nM IC50). C. Effect on Endothelial Cell Growth vs. Proliferation of smooth muscle cells. The promotion of concurrent endothelial cell proliferation with inhibition of smooth muscle cell proliferation is an important consideration for inhibition of exaggerated response to damage caused by vascular reconstruction. The compounds of the present invention increase the growth of human endothelial cells induced by 2% FBS at inhibitory doses to human smooth muscle cells induced by 10% FBS as represented by the sulfated compound of Example 18 as shown in Figure 1. D.

Cytotoxicity:

It was found that cells tolerated very high levels of all compounds very well, however, to ensure that there was no toxicity, the cytotoxicity of the compounds was examined using a commercial modification of the MTT assay (3- [4,5-dimethylthiazol- 2-yl] -2,5-diphenyltetrazolium). Briefly, the cells were re-cultured in 24 well plates to a confluence of 70-80% and, as before, deprived of serum for 24-48 hours prior to the initiation of the experimental protocol. To ensure that the MTT assay monitored toxicity rather than proliferation, cells were incubated with 250 μg of drug in fresh serum-free medium for 24 h. at 37 ° C in a humidified and CO 2 incubator. After consumption of the treatment compound, the indicator dye MTT (3- [4,5-dimethylthiazol-2-yl] -2,5-diphenyltetrazolium bromide) was added for 4 hours at 37 ° C. Cells were then lysed and aliquots from each well were transferred to a 96-well plate for analysis. Absorbance at wavelength 570 nm with a reference wavelength of 630 nm was recorded using an ELISA plate reader. Results are presented as percentage of viable standards without drug use (100% viable) and pre-solubilization (0% viable). No toxicity was observed up to 250 μΜ with the sulfated compounds of Examples 10-25 and Example 26 (Step 5).

Anticoagulant Activity The anticoagulant activity of the compounds of this invention was evaluated in a partial thromboplastin time (APTT) assay using normal human plasma collected from 5 donors using the procedure of Fenichel et. al. (Clin. Chem. 1964, 10, 69). A BBL fibrometer with automatic coagulation timer precession was employed using a 0.3 ml probe. A thromboplastin or partially activated by an ellagic acid was used in these experiments. This reagent is added to citrated human plasma equilibrated at 37øC in the plastic cavity of the coagulation timer. Calcium is added at 37 ° C, the coagulation timer is started, and the fibrin clot formation time (in seconds) is recorded. The effect of compounds added to plasma at concentrations of 12.5-200 pg / mL was determined. Plasmas that did not coagulate after 240 seconds were given a coagulation time of 240 seconds. An unfertilized heparin comparator was used in the concentration range between 1.25 - 10 pg / ml. Coagulation tests at all concentrations were done in triplicate. The analysis of variance of a randomly assigned block was used to determine the significance of the observed differences in coagulation times. Potency is reported in relation to heparin in which the &gt; 1 indicates weaker activity relative to heparin in a pg / ml comparison.

EXAMPLE 25 SULFACTANT COMPOUNDS Antiproliferation of Porcine Smooth Muscle Cells IC 50 or (% Inhibition at Concentration x) Heparin Anticoagulatory Activity (APTT) EXAMPLE 26, STEP 5 118 pM nt EXAMPLE 25 (8% inhibition at 50 pg / mL) nt EXAMPLE 10 161 pM nt-18-

Continuation of Table I Example SULPHATE COMPOUNDS Antiproliferation of Porcine Smooth Muscle Cells IC50 or (% Inhibition at Concentration x) Anticoagulant Activity (ΑΡΤΤ) Relative to Heparin EXAMPLE 24 58 μΜ nt EXAMPLE 11 117 μΜ nt EXAMPLE 12 72 μΜ nt EXAMPLE 13 40.5 - 80 μΜ nt EXAMPLE 14 3 μΜ 6.7 EXAMPLE 15 4.7 μΜ 2.0 - 2.3 EXAMPLE 20 45 μΜ nt EXAMPLE 21 43 μΜ nt EXAMPLE 22 5.2 - 22 μΜ 2.0 EXAMPLE 16 3.6 μΜ 2.2

Continuation of Table I Example SULPHATE COMPOUNDS Antiproliferation of Porcine smooth muscle cells IC50 or (% concentration inhibition x) Anticoagulant activity (APTT) in relation to Heparin EXAMPLE 17 23 - 50.8 μ insign insignificant activity at 50 pg / ml EXAMPLE 18 0.85 μΜ 2.1 EXAMPLE 19 (10-50% inhibition at 20 μΜ) nt EXAMPLE 23 9.5-40 μΜ 2.0 heparin (H-7005) (45-83% inhibition at 50 μg / mL) 1 nt = not tested

Specific procedures are described in the examples which follow. These examples are provided to illustrate the invention and should not be construed as limiting the invention set forth in the appended claims. The terms &quot; Sephadex G-10 &quot;, &quot; Dowex &quot;, &quot; Amberlite IR-20 &quot;, &quot; Sephadex LH-20-100 &quot; and &quot; Sephadex SP-C25 &quot; as used herein are Registered Trade Marks. EXAMPLE 1

To 4.0 g (24 mmol) of 4-methyl-3-nitrobenzyl alcohol and 20.0 g (29 mmol) of 4-methyl-1-nitrobenzene of acetobromo-maltose in 60 mL CH 3 NO 2 were added Hg (CN) 2 6.15 g (24 mmol) and HgBr 2 (6.91 g, 19 mmol). After stirring at room temperature overnight, the reaction was quenched with sat. and stirred for 20 min. The reaction mixture was extracted into CH 2 Cl 2. The organic phase was washed with sat. NaCl, dried (MgSO4), and flash chromatographed &quot; (1: 2 and 1: 1 EtOAc / petroleum ether). Recromatographed using ether / petroleum ether (3: 1, then 4: 1, then 100: 0) gave 7.97 g of the title compound as a colorless solid: 1 H-NMR (CDCl3, 300 MHz) δ 7.92 (s, 1 H), 7.41 (d, 1 H), 7.32 (d, 1 H), 5.42 (d, 1 H), 5.36 (t, 1 H), 5.25 (d, 1 H), 5.06 (t, 1 H), 4.8-5.0 (m, 3 H), 4.65 (d, 1 H), 4.62 (d, 1 H) , 4.54 (dd, 1 H), 4.2-4.3 (m, 2 H), 3.9-4.1 (m, 3 H), 3.65-3.71 (m, 1 H), 2.60 (s, 3 H), 2.16 (s, 3 H), 2.11 (s, 3 H), 2.04 (s, 3 H), 2.03 (s, 6 H), 2.01 (s, 3 H) and 2.00 ppm (s, 3 H).

Step 2 5- (hepta-Q-acetyl-β-D-maltosyloxymethyl-2-methylphenylamine

A mixture of 4.0 g (5.09 mmol) of 5- (hepta-O-acetyl-PD-maltosyloxymethyl) -2-methyl-1-nitrobenzene prepared in Example 1 and 8.00 g (35 mmol ) of SnCl2 · H2 O in 100 mL EtOAc was heated at reflux for 2 hours. The reaction mixture was cooled to room temperature and quenched with sat. After stirring for 15 min, the mixture was diluted with CH2 Cl2 (200 mL) and filtered through &quot; solka floc &quot; (CH2 Cl2). The organic phase was dried (MgSO4) and concentrated. Chromatography &quot; flash &quot; (1: 2, then 1: 1) afforded 3.42 g (89% yield) of the title compound as a colorless foam: 1 H-NMR (CDCl 3, , 300 MHz) δ 7.01 (d, 1 H), 6.63 (s, 1 H), 6.62 (d, 1 H), 5.41 (d, 1 H), 5.39 (M, 2 H), 4.75 (d, 1 H), 4.70 (t, 1 H), 4.20 , 4.51 (d, 1 H), 4.26 (dd, 2 H), 3.95-4.01 (m, 3 H), 3.63-3.67 (s, 6 H), 2.11 (s, 3 H), 2.03 (s, 6 H), 2.00 (s, 3 H), and 1.99 ppm (s, 6 H).

Procedure B: A solution of 31.1 g (39.6 mmol) of 5- (hepta-O-acetyl-PD-maltosyloxymethyl) -2-methyl-1-nitrobenzene prepared in step 1 of Example 1 was hydrogenated at 50 psi over 10% Pd / C (10.0 g) for 1 h. The catalyst was removed by filtration and the filtrate was concentrated to give a white foam. Trituration with water provided 28.0 g (94%) of the title compound as a white solid, m.p. 154-156 ° C. EXAMPLE 2

The title compound was prepared in 47% yield by the procedure of step 1 of Example 1 using 4-methyl-3-methyl-1-nitrobenzene purification was accomplished by flash chromatography (EtOAc / petroleum ether, 1: 2 to 1: 1 to 2: 1): 1 H-NMR (CDCl3, 300 MHz) δ 7.91 (s, 1 H), 7.40 (d, 1 H), 7.32 (d, 1 H), 4.80-5.20 (m, 6 H), 4.40-4.80 , 4 H), 4.36 (dd, 1 H), 4.02-4.13 (m, 2 H), 3.81 (t, 1 H), 3.60-3.68 (m, 2 H), 2.60 (s, 3 H), 2.14 (s, 3 H), 2.08 (s, 3 H), 2.06 (s, 3 H), 2.03 (s, 3 H), 2.02 (s, 3 H), 2.01 (s, 3 H) and 1.98 ppm (s, 3 H)

The title compound, mp 180 DEG-182 DEG C., was prepared in 62% yield from 5- (hepta-O-acetyl-PD-2-propyl-2-methyl- 2-methyl-1-nitrobenzene using Procedure A from step 2 of Example 1. Purification was achieved by &quot; flash chromatography &quot; (DMSO-d6, 400 MHz) δ 6.86 (d, 1 H), 6.48 (d, 1 H), 6 (EtOAc / petroleum ether, 1: 1 to 2: 1) , 36 (dd, 1 H), 5.24 (t, 1 H), 5.10 (m, 1 H), 4.80-4.90 (m, 3 H), 4.61-4.72 (m, 2H), 4.56 (d, 1 H), 4.30-4.31 (m, 2 H), 4.22 (dd, 1 H), 4.08 (dd, 1 H) , 3.99-4.03 (m, 1 H), 3.94 (dd, 1 H), 3.73-3.81 (m, 2 H), 2.10 (s, 3 H), 2. (S, 3 H), 1.98 (s, 3 H), 1.96 (s, 3 H), 1.95 (s, 3 H), 1.95 , 94 (s, 3H) and 1.91 ppm (s, 1H); mass spectrum (+ FAB) m / z 778. Anal. Calc. Calc'd for C 34 H 46 NO 8: C, 53.97; H, 6.13; N, 1.85. Found: C, 53.67; H, 5.92; N, 1.62. EXAMPLE 3

Step 1 5- (Hepta-O-acetyl-BD-cellobiosyloxymethyl) -2-chloro-1-nitrobenzene The title compound was prepared in 45% yield by the procedure described in step 1 of Example 1 using 4-chloro-3- (d, 1 H), 7.42 (dd, 1 H), 4.80 (d, 1 H) 5.20 (m, 5 H), 4.73 (d, 1 H), 4.51-4.66 (m, 4 H), 4.30-4.40 (m, 1 H) (M, 2H), 3.81 (t, 1H), 3.60-3.68 (m, 2H), 2.13 (s, 3H), 2.09 (m, 2.03 (s, 3 H), 2.01 (s, 3 H), 2.01 (s, 3 H) s, 3 H) and 1.57 ppm (s, 3 H)

The title compounds were prepared from 5- (hepta-O-acetyl-PD-cellobiosyloxymethyl) -2-chloro-1-nitrobenzene with methyl 61% yield as a solid by trituration with ether using Procedure A from step 1 of Example 1: 1 H-NMR (CDCl3, 300 MHz) δ 7.20 (d, 1 H), 6.75 (s, 1 H), 6.60 (d, 1 H), 4.89-5.19 (m, 5 H), 4.73 (d, 1 H), 4.47-4.60 (m, 4 H) , 4.36 (dd, 1 H), 4.02-4.13 (m, 2 H), 3.80 (t, 1 H), 3.55-3.67 (m, 2 H), 2. (S, 3 H), 2.03 (s, 3 H), 2.02 (s, 6 H), 2.01 (s, 3 H), and 2 , Δ ppm (s, 3H). EXAMPLE 4

Step 1 2- (Hepta-O-acetyl-BD-cellobiosyloxymethyl) -4-chloro-1-nitrobenzene The title compound was prepared in 47% yield by the procedure of step 1 of Example 1 using 5-chloro-2- nitrobenzoic acid and acetobromocelobiose. Partial purification was achieved using &quot; flash chromatography &quot; (CDCl 3, 300 MHz) δ 8.09 (d, 1 H), 7.71 (d, 1 H), 7.92 (d, 1 H) (Dd, 1 H), 4.90-5.25 (m, 8 H), 4.63 (d, 1 H), 4.52 (d, 1 H), 4.49 (d, 1 H) ), 4.38 (dd, 1 H), 4.03-4.16 (m, 3 H), 3.79 (t, 1 H), 3.60-3.68 (m, 2 H), (S, 6 H), 2.04 (s, 6 H), 2.01 (s, 3 H) and 1.98 ppm (s, 3 H) . The title compound was prepared in 55% yield from 2- (Hepta-O-acetyl-PD-cellobiosyloxymethyl) -5-chloro-1-methyl-2- (hepta-O-acetyl-D-cellobiosyloxymethyl) -4-chlorophenylamine nitrobenzene by Procedure A of step 2 of Example 1. Purification was achieved by trituration with ether: 1 H-NMR (CDCl3, 300 MHz) δ 7.11 (dd, 1H), 7.04 (d, 1H), 6.65 (d, 1 H), 5.03-5.18 (m, 3 H), 4.88-4.97 (m, 2 H), 4.45-4.80 (m, 5 H ), 4.36 (dd, 1 H), 4.02-4.13 (m, 2 H), 3.60-3.81 (m, 3 H), 2.16 (s, 3 H), 2.08 (s, 3H), 2.08 (s, 3H), 2.04 (s, 3H), 2.01 (s, 6H) EXAMPLE 5

Step 1 The title compound was prepared in 54% yield by the procedure described in step 1 of Example 1 using .alpha.-dodecylbromide tetraacetate, D-glucopyranosyl and 4-methyl-3-nitrobenzyl alcohol: ¹H-NMR (CDCl,, 300 MHz) δ 7.92 (d, 1H), 7.43 (dd, 1H), 7.33 (d, (D, 1 H), 4.66 (d, 1 H), 4.56 (d, 1 H) (S, 3 H), 2.11 (s, 3 H), 2.65 (s, 3 H) ), 2.07 (s, 3H), 2.03 (s, 3H) and 2.01 ppm (s, 3H).

Step 2 5- (Tetra-O-acetyl-BD-glucopyranosyloxymethyl) -2-methylphenylamine The title compound was prepared in 63% yield by Procedure A from step 2 of Example 1 from 5- (tetrahydrofuran- -acetyl-PD-glucopyranosyloxymethyl) -2-methyl-1-nitrobenzene and purified by trituration of the crude reaction mixture with ether: 1 H NMR (CDCl3, 300 MHz) δ 7.02 (d, 1H), 6.64 (s, 1 H), 6.63 (d, 1 H), 5.02-5.17 (m, 3 H), 4.79 (d, 1 H), 4.50-4.55 (M, 1 H), 2.18 (s, 3 H), 2.17 (dd, 1 H) , 11 (s, 3 H), 2.02 (s, 3 H), 2.01 (s, 3 H) and 2.00 ppm (s, 3 H).

The title compound was prepared in 42% yield by the procedure described in step 1 of Example 1 using an equivalent of 5-nitro- (2-chlorobenzyloxy) -1- (EtOAc / CH 2 Cl 2, 3: 1) and trituration with ether: 1 H NMR (DMSO-d 6):? NMR (CDCl3, 300 MHz) δ 8.10 (s, 2 H), 7.48 (s, 1 H), 4.88-5.30 (m 12 H), 4.68 (d, 2 H) ), 4.38 (dd, 2 H), 4.00-4.13 (m, 4 H), 3.82 (t, 2 H), 2.08 (s, 6 H), 2.07 (s, 6 H), 2.03 (s, 6 H), 2.08 12 H), 2.01 (s, 6 H) and 1.98 ppm (s, 6 H).

Step 2 3,5-Bis (hepta-O-acetyl-BD-cellobiosyloxymethyl) phenylamine The title compound was prepared in 54% yield by Procedure A from step 2 of Example 1 using 3,5-bis (hepta-O- acetyl-β-D-cellobiosyloxymethyl) -1-nitrobenzene. Purification was achieved by &quot; flash chromatography &quot; (EtOAc / petroleum ether, 1: 1): 1 H-NMR (CDCl3, 300 MHz) δ 6.73 (bs, 2 H), 6.67 (bs, 1 H), 5.03-5 (M, 6H), 4.97 (d, 2H), 4.91 (d, 2H), 4.75 (d, 2H), 4.49-4.61 (m, 6H). H), 4.37 (dd, 2 H), 4.02-4.13 (m, 6 H), 3.81 (t, 2 H), 3.57-3.69 (m, 4 H) , 2.15 (s, 6 H), 2.01 (s, 6 H), 2.08 (s, 6 H), 2.03 ). EXAMPLE 7

The title compound was prepared by the procedure described in step 1 of Example 1 using an equivalent of 5-nitro-m-xylene-1,2,3,4-tetrahydronaphthalen- Purification was achieved initially by flash chromatography (EtOAC / CH 2 Cl 2, (3: 1)) and then rechromatography first with toluene / EtOAc (1: 1) and then with EtOAc / CH 2 Cl 2 (1: 4 to 1: 2) to afford the partially purified product which was used directly in the next reaction

The title compound was prepared by procedure A from step 2 of Example 1 using 3,5-bis (hepta-O-acetyl-PD-cellobiosyloxymethyl) -1-hydroxy- nitrobenzene Partial purification was achieved by flash chromatography (CH2 Cl2 / EtOAc 2: 1 to 1: 2)) and then rechromatographed (CH 2 Cl 2 / EtOAc 3: 1 to 1: 1)) to give the title compound. EXAMPLE 8

Step 1 3,5-Bis (tetra-O-acetyl-BD-glucosyloxymethyl) -nitrobenzene The title compound was prepared by the procedure described in step 1 of Example 1 using an equivalent of 5-nitro-m-xylene- Purification was achieved by flash chromatography (EtOAc / CH 2 Cl 2, (1: 4 to 1: 2)) to give 29% yield of the title compound as a white solid. almost pure title which was used without further purification.

Step 2 3,5-Bis (tetra-O-acetyl-BD-glucosyloxymethyl) phenylamine The title compound was prepared by Procedure A from Step 2 of Example 1 using 6.27 g of 3,5-bis (tetra-O-acetyl- BD-glucosyloxy-methyl) -1-nitrobenzene. Purification was achieved by &quot; flash chromatography &quot; (CH 2 Cl 2 / EtOAc (3: 2)) to give 2.00 g (39% yield) of the title compound as a yellow oil: partial δ-δ (CDCl3, 300 MHz) δ 6.56 (s, 3 H), 5.0-5.2 (m, 6 H), 4.8 (d, 2 H), 4.4-4.6 (m, 4 H), 3.7 ppm H).

The title compound was prepared by the procedure described in step 1 of Example 1 using 2.57 g (14.02 mmol, 1 equivalent) of the title compound as a white solid 5-nitro-m-xylene-α, α'-diol, two equivalents of acetobromomaltose and two equivalents of all other reagents, all in CH 3 NO 2 (200 mL) Purification was achieved by flash chromatography (EtOAc / Recromatography (Et2 O) gave 11.4 g (57% yield) of the title compound which was used in the following reaction: 1 H-NMR (CDCl3, 300 MHz) Δ 8.11 (s, 2H), 7.49 (s, 1H), 5.42 (d2H), 5.36 (t, 2H), 5.26 (t, 2H) , Δ (t, 2H), 4.21-4.29 (m, 4H) and 3.69-3.72ppm (m, 2H).

The title compound was prepared by procedure A from step 2 of Example 1 using 3,5-bis (hepta-O-acetyl-β-D-maltosyloxymethyl) - (CH 2 Cl 2 / EtOAc, (1: 1)) to afford 4.43 g (40% yield) of the title compound: 1 H-NMR (CDCl 3, 300 MHz) δ 6.57 (s, 3 H), 5.41 (d, 2 H), 5.36 (t, 2 H), 5.23 (t, 2 H), 5.50 (t, 2 H), 4.75 (d, 2 H), 4.26 (m, 4 H) and 3.65-3.69 ppm (m, 2 H) EXAMPLE 10

Step 1

Dodecanedioic acid bis (β-D-cellobiosyloxymethyl-2-methylphenylamide)

To a solution of 5- (Hepta-O-acetyl-β-D-cellobiosyloxymethyl) -2-methylphenylamine (1.03 g, 1.36 mmol) and triethylamine (189 μl, 1.36 mmol) in THF

(15 mL) was added dodecanedioyl dichloride (170 æl, 0.681 mmol). After stirring at room temperature for 2 h, the reaction mixture was quenched with MeOH, diluted with CH 2 Cl 2 and then washed with water. The organic phase was dried (MgSO4), filtered and concentrated to give 1.157 g of a colorless solid. This product was dissolved in MeOH (25 mL) and treated with 11.9 mL (11.9 mmol) 1 N NaOH. After stirring at 50 ° C for 5 h, the reaction mixture was cooled to ambient temperature and treated with 9.5 mL (9.5 mmol) 1 N HCl. Collection of the colorless solid gave the title compound (566 mg, 75% yield), mp> 200 ° C: 1 H-NMR (DMSO-d 6, 400 MHz) δ 9.25 (s, 2 H), 7.32 (s, 2 H), 7.16 (d, 2 H) 1 H), 4.77 (d, 2 H), 4.50 (d, 2 H), 4.24-4.30 (two doublets, 2 H), 3.78 (d, 2 H) H), 3.69 (d, 2 H), 3.63 (broad d, 2 H), 2.99 (t, 2 H), 2.30 (t, 4 H), 1.58 , 4 H) and 1.29 ppm (broad s, 12 H); 13 C NMR (DMSO-d 6, 100 MHz) δ 171.1, 136.2, 135.5, 131.1, 130.0, 124.8, 103.2, 101.7, 80.6, 76.8 , 76.4, 75.0, 74.9, 73.3, 73.1, 70.0, 69.5, 61.0, 60.4, 35.7, 28.9, 28.8, 28 , 7, 25.3 and 17.6 ppm; mass spectrum (-FAB) m / z 1115 (M-H); IR (KBr) 3430 and 1660 cm -1. Anal. Calc. Calc'd for C 52 H 20 NO 2 204 • 4 H 2 O: C, 52.51; H, 7.46; N, 2.36. Found: C, 52.53; H, 7.28; N, 2.27.

Step 2

Dodecanedioic acid bis (2-methyl-5- (hepta-β-sulfato-β-D-cellobiosyloxymethyl) phenyl]

A solution of the dodecanoic acid bis {[5- (PD-cellobiosyloxymethyl) -2-methylphenyl] amide (391 mg, 0.350 mmol) and triethylamine sulfur trioxide complex (3.66 g, 26.3 mmol) in DMF (40 mL) was heated at 70 ° C for 3 days. The reaction mixture was concentrated in vacuo and passed through a Sephadex G-10 column (twice). Cation exchange was performed using a strongly acidic Dowex 50 x 8 column (Na-form) to give 433 mg of the title compound: 1 H-NMR partial (D2 O, 400 MHz) δ 7.39 (d, 2H ), 4.36 (d, 2H), 7.29 (s, 2H), 4.94 (d, 2H), 4.80-4.90 (m, 4H), 4.59 (d, dd, 2H), 2.49 (t, 4H), 2.23 (s, 3H), 2.30-2.40 (m, 4H) and 1.30-1.50 ppm , 12H); 13 C NMR (D2O, 100 MHz) δ 176.8, 135.2, 134.9, 134.4, 130.9, 127.8, 127.1, 100.0, 99.1, 77.7, 77 , 4.77,1.74.4, 73.7, 73.5, 73.1, 70.7, 67.7, 66.6, 35.8, 28.5, 28.3, 25.5 and 16.9 ppm; mass spectrum (&quot; electrospray &quot; negative) (m-zNa) z 825.6 (M-3Na) 3 ', 613.5 (M-4Na) 4' and 486.2 (M-5Na) 5 '. Anal. Calc. for C52 H66 N2 O6 Si4 Na14.14 H2 O: C, 18.83; H, 2.70; N, 0.80; S, 17.40. Found: C, 17.96; H, 2.57; N, 0.72; S, 17.11. EXAMPLE 11

To 813 mg (1.08 mmol) of 5- (hepta-O-acetyl-PD-cellobiosyloxymethyl) -2-methylphenylamine in THF (20 ml) was added N-bis-5- (B-cellobiosyloxymethyl) -2-methylphenyl] mL) containing triethylamine (148 æl, 1.08 mmol) was added terephthaloyl chloride (109 mg, 0.538 mmol). After stirring at room temperature for 2 h, the reaction mixture was quenched with MeOH, diluted with CH 2 Cl 2 and washed Drying (MgSO4) and concentration gave the crude product, which was diluted with MeOH (25 mL) and treated with 8.7 mL (8.7 mmol) 1 N NaOH. After stirring at 50 ° C for 3 h, the suspension was filtered to give 512 mg (89%) yield of the title compound as a colorless solid, mp> 210 ° C: 1 H-NMR (DMSO-d6, 400 MHz) δ 10.07 (s, 2H), 8.10 (s, 4H), 7.35 (s, 2H), 7.26 (d, 2H), 7.22 (d, 2H), 5.22 -5.24 (m, 4 H), 5.01 (d, 2 H), 4.99 (d, 2 H), 4.82 (d, 2 H), 4.68 (d, 2 H) , 4.54-4.64 (m, 6 H), 4.32 (d, 2 H), 4.25 (d, 2 H), 3.75-3.80 (d, 2 H), 3. , 63-3.77 (m, 4 H), 3.37-3.41 (m, 2H), 2.97-3.19 (m, 10 H) and 2.23 ppm (s, 6 H); 13 C NMR (D2O, 100 MHz) δ 164.6, 137.0, 136.0, 135.9, 133.0, 130.1, 127.7, 126.0, 125.6, 103.2, 101 , S, 80.6, 76.8, 76.4, 75.1, 74.9, 73.3, 73.2, 70.0, 69.3, 61.0, 60.4 and 17.7 ppm; mass spectrum m / z 1052, 889 and 725. Anal. Calc. Calc'd for C 48 H 64 N 2 O 4: C, 54.75; H, 6.13; N, 2.66. Found: C, 55.54; H, 6.33; N, 2.57.

Step 2

The tetradecassodic salt of N, N'-bis-S-diepta-O-sulfato-β-D-cellobiosyloxyethyl) -2-methylphenyl ether tetallamide

A mixture of N, N'-bis [5- (β-cellobiosyloxymethyl) -2-methylphenyl] -tertallamide (321 mg, 0.305 mmol) and trimetylamine sulfur trioxide complex (3.06 g, 22.0 mmol) in DMF (25 mL) was stirred at 70 ° C for 4 days. The reaction mixture was concentrated and passed through a Sephadex G-10 column. 1 H-NMR analysis showed incomplete sulfation. The product was re-subjected to sulfation with 3.06 g (22.0 mmol) of trimethylamine sulfur trioxide complex in 25 mL of DMF at 70 ° C for 5 days. The reaction mixture was cooled to room temperature, concentrated in vacuo and passed through a Sephadex G-10 column. Cation exchange using a Dowex (50 x 8, strongly acidic, Na-form) column afforded 469 mg (62% yield) of the title compound, mp 178øC (dec.): 1 H-NMR (D2 O, 400 MHz) δ 8.11 (s, 4 H), 7.40-7.50 (m, 6 H), 4.98 (d, 2 H), 4.95 (d, 2 H) 32 ppm (s, 6 H); 13 C NMR (D2 O, 100 MHz) δ 169.5, 136.9, 135.4, 135.2, 134.3, 131.0, 128.1, 128.0, 127.0, 100.0, 99 , 77.7, 77.4, 77.3, 77.1, 74.4, 73.7, 73.4, 73.1, 70.7, 67.7, 66.7 and 16.8 ppm. Anal. Calc. Calc'd for C 40 H 50 N 2 O 6 S 6 N 4 O 4 úH 2 O: C, 20.58; H, 2.51; N, 1.00; S, 17.17. Found: C, 20.21; H, 2.84; N, 0.96; S, 17.34. EXAMPLE 12

Step 1

To a solution of 887 mg (1.18 mmol) of 5- (hepta-O-acetyl-PD-cellobiosyloxymethyl) -2-methylphenylamine bis (β- (β-cellobiosyloxymethyl) -amide- and 162 μL (1.18 mmol) triethylamine in THF (20 mL) was added 164 mg (0.588 mmol) biphenyl-4,4'-carboxylic acid chloride. After stirring at room temperature for 4 h, the mixture The organic phase was dried (MgSO4), filtered and concentrated to a colorless solid (1.01 g). A solution of the crude material (985 mg, 0.574 mmol) in methanol MeOH (25 mL) containing 9.18 mL (9.18 mmol) 1 N NaOH was stirred for 3 h at 50 DEG C. The reaction mixture was cooled, and the title compound was collected as a white powder (444 mg, 69% yield): 1 H-NMR (DMSO-d 6, 300 MHz) δ 10.01 (s, 2 H), 8.13 (d, 4 H), 7.94 (d, 4 H), 7.37 (s, 2 H), 7.28 (d, 2 H), 7.23 (d, 2 H), 5.24 (d, 2 H), 5.00 (q, 2 H), 4.84 (d, 2H), 4.56-4 , 69 (m, 4 H), 4.33 (d, 2 H), 4.27 (d, 2 H) and 2.25 ppm (s, 3 H).

Step 2

Tetradecassic salt of biphenyl-4,4'-dicarboxylic acid bis (5-hepta-β-sulfato-β-D-cellobiosyloxymethyl) -2-

A solution of biphenyl-4,4'-dicarboxylic acid bis {[5- (PD-cellobiosyl-xymethyl) -2-methylphenyl] amide 437 mg (0.387 mmol) and 3.85 g (27.7 mmol) of trimethylamine sulfur trioxide complex in DMF (25 mL) was stirred at 70 ° C for 5 days. The reaction mixture was quenched with water and concentrated in vacuo. The residue was dissolved in a small amount of water and passed through a Sephadex G-10 column. Exchange of cations was achieved using strongly acidic resin (Na-form) Dowex 50 x 8 to afford, after azeotroping with toluene, 742 mg (75% yield) of the title compound, mp 170øC (dec.): (D, 4 H), 7.97 (d, 4 H), 7.40 (m, 6 H), 4.98 (d, 2 H), 4.94 (d, 2H), 4.63-4.68 (m, 4 H), 4.56 (dd, 2 H), 4.41 (dd, 2 H), 4.30-4, 37 (m, 6 H), 4.17-4.23 (m, 4 H), 3.99-4.05 (m, 4 H) and 2.31 ppm (s, 6 H); 13 C NMR (D2O, 100 MHz) δ 169.7, 143.2, 135.3, 135.2, 134.4, 132.8, 130.9, 128.1, 128.0, 127.4, 127 , 99.9, 98.9, 77.6, 77.3, 77.2, 77.0, 74.2, 73.6, 73.3, 73.0, 70.6, 67.6 , 66.6 and 16.7 ppm. Anal. Calc. Calc'd for C 55 H 54 N 2 O 6 Si 4 Na 14.14 H 2 O: C, 21.97; H, 2.80; N, 0.95; S, 16.29. Found: C, 21.77; H, 2.90; N, 0.95; S, 13.66. EXAMPLE 13

To a solution of 887 mg (1.18 mmol) of 5- (hepta-O-acetyl-PD-cellobiosyloxymethyl) -2-methylphenylamine in THF (1: 1) in dichloromethane 20 mL) containing triethylamine (162 μl, 1.18 mmol) was added isophthaloyl dichloride (119 mg, 0.587 mmol). After stirring at room temperature for 2 h, the reaction mixture was quenched with MeOH, diluted with CH 2 Cl 2 and washed with water. The organic phase was dried (MgSO4) and concentrated to give 1.00 g of a colorless solid. A solution of the crude product in MeOH (25 mL) containing 9.76 mL (9.76 mmol) 1 N NaOH was stirred at 50 ° C for 3 h. The resulting solid was collected and dried azeotroped over toluene to provide 355 mg (55% yield) of the title compound as a colorless powder, mp 200-203øC: 1 H-NMR (DMSO-d 6, 400 MHz ) δ 10.09 (s, 2 H), 8.54 (s, 1 H), 8.15 (d, 2 H), 7.68 (t, 1 H), 7.34 (s, 2 H ), 7.26 (d, 2H), 7.21 (d, 2H), 5.24 (t, 4H), 5.01 (dd, 2H), 4.82 (d, 2H , 4.69 (s, 2H), 4.54-4.69 (m, 4 H), 4.32 (d, 2H), 4.25 (d, 2H), 3.76- 3.80 (m, 2 H), 3.63 - 3.69 (m, 4 H) and 2.24 ppm (s, 6 H); 13 C NMR (DMSO-d 6, 100 MHz) δ 164.9, 136.1, 135.9, 134.8, 133.0, 130.5, 130.1, 128.6, 127.1, 125.9, 125.6, 103.2, 101.8, 80.6, 76.8, 76.4, 75.1, 74.9, 73.3, 73.2, 70.0, 69.4, 61, 0, 60.4 and 17.7 ppm; mass spectrum (-FAB) m / z 1051, 727 and 889. Anal. Calc. Calc'd for C 48 H 64 N 2 O 243 H 2 O: C, 52.08; H, 6.37; N, 2.53. Found: C, 51.89; H, 6.31; N, 2.26.

Step 2

N-Bis-S-fhepta-O-suifate-S-D-cellobiosyloxymethyl-2-methylphenylisophthalamide

A solution of N, N'-bis [5- (PD-cellobiosyloxymethyl) -2-methylphenyl] isophthalamide (180 mg, 0.171 mmol) and 1.79 g (11.98 mmol) of trimethylamine sulfur trioxide complex in DMF (25 mL) was stirred at 70 ° C for 4 days. The reaction mixture was concentrated and then purified using a Sephadex G-10 column eluting with water. Cation exchange was performed with a Dowex 50 x 8 column of strongly acidic resin (Na-form) eluting with water. Removal of the water in vacuo and azeotroping with toluene gave 358 mg (84% yield) of the title compound as a colorless solid, mp 171øC (dec.): 1 H-NMR (D2 O, 400 MHz) δ 8.43 (s, 2 H), 8.21 (dd, 2 H), 7.78 (t, 1 H), 7.40-7.50 (m, 6 H), 4.97 , 2 H), 4.94 (d, 2 H), 4.56 (dd, 2 H) and 2.30 ppm (s, 6 H); 13 C NMR (D2O, 100 MHz) δ 169.2, 135.3, 135.1, 134.3, 134.0, 131.0, 130.9, 129.3, 128.0, 126.9, 126 , 5.99,9.99, 135.5, 77.3, 77.2, 77.0, 74.3, 73.6, 73.3, 73.0, 70.6, 67.6 , 66.6 and 16.7 ppm. Anal. Calc. Anal. Calc'd for C 48 H 20 N 2 O 6S 6 4 Si 144 H 2 Na 2 SO 4: C, 19.12; H, 2.61; N, 0.93; S, 17.01. Found: C, 18.97; H, 2.45; N, 0.94; S, 15.68. EXAMPLE 14

Step 1

To a solution of 3,5-bis (hepta-O-acetyl-PD-cellobiosyloxymethyl) -phenylamine (817 mg, 0.588 mmol) and triethylamine (82 ÂμL, 0.59 mmol) in THF (25 mL) was added 74 μl (0.295 mmol) of dodecanedioyl dichloride. The reaction mixture was stirred at room temperature for 90 min, quenched with MeOH, diluted with CH 2 Cl 2 and washed with water. The organic phase was dried (MgSO4) and concentrated to a colorless solid (834 mg, 95% yield). The crude product was dissolved in MeOH (25 mL) and treated with 8.4 mL (8.4 mmol) of 1 N NaOH. After stirring at 50 ° C for 2 h, the reaction mixture was quenched at ambient temperature with 7.84 mL of 1N HCl, concentrated and purified by reverse phase column chromatography (silica RP 60) using MeOH / Recromatography using MeOH / H2 O (2: 3) gave 438 mg (87% yield) of the title compound as a colorless solid: 1 H-NMR (D2 O, 400 MHz) δ 7.45 (s, 4 H), 7.26 (s, 2 H), 4.64 (d, 4 H), 4.47 (t, 8 H), 3.80 (dd, 4H), 3.71 (dd, 4H), 2.30 (broad t, 4 H), 1.55 (t, 4 H) and 1.17 ppm (broad m, 12 H); 13 C NMR (D2O, 100 MHz) δ 175.1, 138.1, 137.7, 124.4, 120.3, 102.5, 101.3, 78.7, 75.9, 75.5, 74 , 7.74, 73.3, 73.1, 72.8, 70.8, 69.4, 60.5, 60.0, 36.6, 28.7, 28.6, 28.5 and 25.3 ppm. Anal. Calc. for CHHH20HH₂46O 10: 10 H₂O: C, 46.15; H, 7.13; N, 1.42. Found: C, 45.91; H, 6.82; N, 1.41. GgfBSSSSS ^

Step 2

Bis (r3 <5-bis (hepta-β-sulfato-β-d-cellobiosyloxymethyl) -dimino) decane

A solution of the decaniodic acid bis {[3,5-bis (BD-cellobiosyloxymethyl) phenyl] amide} (177 mg, 98.4 mmol) and trimethylamine sulfur trioxide complex (1.97 g, 14.2 mmol) in DMF (25 mL) was stirred at 70 ° C for 25 h. The reaction mixture was concentrated and purified by Sephadex G-10 chromatography using water to elute. Cation exchange using a Dowex 50 x 8 column of strongly acidic resin (Na-form) afforded 292 mg (64% yield) after azeotroping with toluene: Partial1 H-NMR (D2 O, 400 MHz) δ 7, (S, 4 H), 7.35 (s, 2 H), 4.96-4.98 (m, 8 H), 2.45 (bt, 4 H), 1.40-1.60 (bt, 4 H) and 1.36-1.41 ppm (broad m, 12 H); , Δ 176.1, 138.0, 137.1, 124.5, 121.0, 99.9, 99.6, 77.6, 77.43, 77.39, 77.0, 74.3, 73.4, 72.9, 70.9, 67.6, 66.3, 36.5, 28.7, 28.5 and 25.3 ppm. Anal. Calc. Calc'd for C 76 H 92 N 2 O 3 • 3 N 2 SO 4 • 28H 2 O: C, 16.34; H, 2.67; N, 0.50; S, 17.8. Found: C, 16.25; H, 2.63; N, 0.59; S, 17.07. EXAMPLE 15

Step 1

To a solution of 3,5-Bis (hepta-O-acetyl-PD-cellobiosyloxymethyl) phenylamine (817 mg, 0.588 mmol) and triethylamine (86 mg, 0.588 mmol) in dichloromethane. (86 mg, 0.309 mmol) After stirring at room temperature for 2 h, the reaction mixture was quenched with dichloromethane MeOH, diluted with CH2 Cl2 and washed with H2 O. The reaction mixture was dried (MgSO4) and concentrated to give 921 mg of the crude product, which was dissolved in MeOH (20 mL) and treated with 9.3 mL of 1 N NaOH. After stirring at 50 ° C for 4 h, 1 N HCl (8.6 mL, 8.6 mmol) was added to the cold reaction. Recovering the solid provided 482 mg (86% yield) of the title compound: 1 H (DMSO-d6, 400 MHz) δ 10.40 (s, 2 H), 8.11 (d, 4 H), 7.93 (d, 4 H), 7.74 (s, 4 H (D, 4 H), 4.56 (d, 4 H), 4.35 (d, 4 H), 4.27 (d, 4 H) ), 3.80 (d, 4 H), and 2.97-3.07 ppm (m, 4 H); 13 C NMR (DMSO-d 6, 100 MHz) δ 165.0, 142.0, 138.9, 138.2, 134.1, 128.5, 126.9, 122.8, 119.3, 103.2, 102.0, 80.5, 76.8, 76.5, 75.1, 75.0, 73.3, 73.2, 70.02, 69.96, 61, 0 and 60.4 ppm; IR (KBr) 1650 cm -1. Anal. Calc. Calc'd for C78 H18 N2 O4: 0.4H2 O: C, 47.08; H, 6.48; N, 1.41. Found: C, 46.64; H, 6.22; N, 1.62.

Step 2

Octacosodical salt of biphenyl-3,5-bis (hepta-β-sulfato-β-D-cellobiosyloxymethyl)

A solution of 338 mg (0.187 mmol) of biphenyl-4,4'-dicarboxylic acid bis {[3,5-bis (PD-cellobiosyloxymethyl) phenyl] amide} and 3.64 g (26.1 mmol) of triethylamine sulfur trioxide in DMF (25 mL) was stirred at 70 ° C for 2 days. The reaction mixture was cooled to room temperature, quenched with water and concentrated. Purification was achieved with a Sephadex G-10 column (elution with H2 O). Cation exchange was effected by passing an aqueous solution of the product through a Dowex 50 x 8 column with strongly acidic resin (Na-form). Removal of the solvent and azeotroping with toluene gave 765 mg (88% yield) of the compound as an off-white solid, mp 180 ° C (dec): Partial1 H-NMR (D2 O, 400 MHz) δ 8 , 7.97 (d, 4 H), 7.70 (s, 4 H), 7.39 (s, 2 H), 5.02 (d, 4 H), 4 , 98 (d, 4 H), 4.15-4.23 (m, 8 H) and 4.00 ppm (m, 8 H); , Δ 169.2, 143.3, 138.2, 137.3, 133.5, 128.3, 127.5, 125.0, 121.8, 100.0, 99.7, 77.7, 77.54, 77.51, 77.1, 74.4, 73.5, 73.0, 71.0, 67.7 and 66.4 ppm; mass spectrum (&quot; electrospray &quot;) (m-zNa) / z 401.3 (m-11 Na) 11 ', 443.7 (m -ONa) 10', 495.6 (m- , 560.4 (m-8 Na) 8 'and 643.7 (m-7 Na) 7'.

Anal. Calc. Calc'd for C 78 H 80 N 2 O 13 S 28 Na 28 28 Na 2 SO 4 ú28 H 2 O: C, 17.17; H, 2.51; N, 0.51; S, 16.46. Found: C, 17.06; H, 2.11; N, 0.55; S, 12.04. EXAMPLE 16

Step 1

To a solution of 3,5-bis (hepta-O-acetyl-D-lactosyloxymethyl) -3,5-bis (hepta-O-acetyl- (D-lactosyloxymethyl) phenyl] amide of biphenyl-4,4,4- phenyl-amine (1.05 g, 0.757 mmol) and triethylamine (105 æL, 0.757 mmol) in THF (20 mL) was added 4,4-biphenyldicarboxylic acid dichloride (106 mg, 0.378 mmol). The reaction mixture was dried (MgSO4), concentrated and purified by trituration with CH2 Cl2 / Et2 O to give 984 mg (87% yield) of the title compound as a colorless oil. title compound as a colorless solid, mp 196-167øC: 1 H NMR (CDCl3, 400 MHz) δ 8.62 (s, 2H), 8.05 (d, 4 H), 7.74 ( d, 4 H), 7.66 (s, 4 H), 6.93 (s, 2 H), 5.33 (d, 2 H), 5.17 (t, 2 H), 5.07- (M, 4 H), 4.79 (d, 2 H), 4.62-4.66 (m, 4 H) (D, 2H), 4.03-4.15 (m, 6 H), 3.80-3.88 (m, 4 H), 3.62 (d, 2 H) d, 2H), 2.13 (s, 12H), 2.10 (s, 12H), 2.03 (s, 24 H), 2.02 (12 H) and 1.95 ppm (s, 12 H); mass spectrum (&quot; electrospray &quot;, Ca 2+ adduct) m / z 1513.2 (m + Ca) 2+. Anal. Calc. for C134H164N2O741H2O; C, 53.56; H, 5.57; N, 0.93. Found; C, 53.25; H, 5.55; N, 1.06.

Step 2

Bis-biphenyl-O-silanyloate-B-D-lactosyloxymethyl) phenylamide) biphenyl-4-dicarboxylic acid octa-sodium salt

A solution of 871 mg (0.292 mmol) of biphenyl-4,4'-dicarboxylic acid bis {[3,5-bis (hepta-O-acetyl-D-lactosyloxymethyl) phenyl] amide} in MeOH (15 mL) containing 8.75 mL 1 N NaOH (8.75 mmol) was stirred at 50 ° C for 3 h. The reaction mixture was cooled to room temperature and quenched with 1 N HCl (8.16 mL, 8.16 mmol). The resulting solid was azeotroped with toluene: 13 C NMR (DMSO-d6, 190 MHz) δ 164.9, 141.9, 138.9, 138.2, 134.1, 128.4, 126.8, 119.2, 103.8, 101.9, 80.7, 75.5, 99, 74.96, 73.3, 73.2, 70.6, 69.9, 68.0, 60.5 and 60.3 ppm.

A solution of biphenyl-4,4'-dicarboxylic acid bis {[3,5-bis (hepta-O-acetyl-β-D-lactosyloxymethyl) phenyl] amide 396 mg (0.219 mmol) and 4.26 g (30.6 mmol) of sulfur trioxide complex and trimethylamine in DMF (20 mL) was stirred at 70 ° C for 3 days. The reaction mixture was cooled to room temperature, quenched with distilled H2 O and concentrated. The residue was purified by passage through a Sephadex G-10 column (elution with H 2 O). Exchange of cations was achieved by passing an aqueous solution of the product through a Dowex 50 x 8 column of strongly acidic resin (Na-form). Removal of the solvent and azeotroping with toluene gave 664 mg (65%) of a bright white solid, mp 149øC (dec.): 13C NMR (D2 O, 100 MHz) δ 169.1, -40-75.8 , 75.3, 75.05, 75.00, 73.0, 71.1 and 66.34 ppm; mass spectrum (&quot; electrospray &quot;) (m-zNa) / z 643.7 (m-7 Na) 7 ', 754.8 (m- 6 Na) 6' and 910.4 (m- '. Anal. Calc. Calc'd for C 78 H 8 NO 20 13 8 S 28 Na 28 28 H 2 O: C, 18.12; H, 2.65; N, 0.54; S, 17.36. Found: C, 18.06; H, 2.67; N, 0.50; S, 14.75. (NMR and capillary electrophoresis showed a major component and several other minor components; mass spectral analysis indicates products of 20-28 sulfates). EXAMPLE 17

Step 1

To a solution of 3,5-bis (tetra-O-acetyl-PD-glucosyloxymethyl) phenylamine (996 mg, 0.8 mmol) in dichloromethane , 1.22 mmol) and triethylamine (160 æL, 1.22 mmol) in THF (15 mL) was added 4,4-biphenyldicarboxylic acid dichloride (171 mg, 0.612 mmol) After stirring at room temperature for 3 h , the reaction mixture was quenched with MeOH, diluted with CH 2 Cl 2 and washed with H 2 O. The reaction mixture was dried (MgSO 4), concentrated and purified by trituration with CH 2 Cl 2 / Et 2 O to give 984 mg (87% yield) of the title compound. (CDCl3, 400 MHz) δ 8.42 (s, 12 H), 8.04 (d, 4 H), 7.75 (d, 4 H), 4.66 (s, 4 H), 6.98 (s, 2 H), 5.03-5.22 (m, 12 H), 4.85 (d, 4 H) (D, 4 H), 2.06 (s, 12 H), 4.61 (d, 4 H), 4.22-4.32 (m, 8 H) ), 2.024 (s, 12 H), 2.019 (s, 12 H) and 1.99 ppm (s, 12 H) mass spectrum ((-) - FAB), m / z 1159 (MH). Calc for C5 4.20H2 O: C, 51.10; H, 6.35; N, 2.21. Found: C, 51.01; H, 6.09; N, 2.25. -41-

Step 2 Bis (β-D-glucosyloxymethyl) phenyl] amide (b3,5-bis (β-D-glucosyloxymethyl) phenylamide)

A solution of 575 mg (0.495 mmol) of biphenyl-4,4'-dicarboxylic acid bis {[3,5-bis (tetra-O-acetyl-BD-glucosyloxymethyl) phenyl] amide} and 8.91 mL , 9 mmol) of 1N NaOH in MeOH (15 mL) was stirred at 50 ° C for 4 hours. The reaction mixture was quenched with 1N HCl (7.9 mL) and purified by chromatography on Saphadex G-10 (elution with H2 O). Removal of H2 O in vacuo gave 144 mg (25% yield) of the title compound, mp 160øC (dec.): Δ-D (400 MHz, DMSO): δ 7.38 (d, 4 H), 7.20 (s, 4 H), 7.16 (d, 4 H), 7.07 (s, 2 H), 4.72 (d, 4 H), 4.47 (d, 4 H), 4.38 (d, 4 H), 3.88 (d, 4 H), 3.69 (dd, 4 H) and 3.27 - 3.44 ppm (m, 16 H); 13 C NMR (D2 O, 100 MHz) δ 166.3, 141.6, 137.6, 137.2, 131.7, 127.5, 126.3, 124.0, 120.1, 101.5, 75 , 7, 75.6, 73.0, 70.7, 69.5 and 60.6 ppm; mass spectrum ((-) - FAB), m / z 1159.4 (M-H), 997.3, 981.3 and 699.2. Anal. Calc. Calc'd for C54 H68 N2 O6 • 6H2 O: C, 51.10; H, 6.35; N, 2.21. Found: C, 51.01; H, 6.09; N, 2.25.

Step 3

Hexadecodicarboxylic acid bis (3,5-bis (tetra-β-sulfato-β-D-glucosyloxymethylphenylamide) hexadecanoic acid salt of biphenyl) -dicarboxylic acid

A solution of 92 mg (0.079 mmol) of biphenyl-4,4'-dicarboxylic acid bis {[3,5-bis (p-D-glucosyloxymethyl) phenyl] amide} and 450 mg (6.83 mmol) of trioxide complex of sulfur and trimethylamine in DMF (15 mL) was stirred at 70 ° C for 3 days. The reaction mixture was cooled to room temperature, quenched with H2 O, distilled and concentrated. The residue was purified by passage through a Sephadex G-10 column (elution with H 2 O). Exchange of cations was achieved by passing an aqueous solution of the product through a Dowex 50 x 8 column of strongly acidic resin (Na-form). Removal of the solvent and azeotroping with toluene gave 161 mg (73%) of a bright white solid, mp 172øC (dec.): 1 H-NMR (D2O, 400 MHz) δ 8.10 (dd, 4 H) , 7.97 (dd, 4 H), 7.71 (s, 4 H), 7.40 (s, 2 H), 5.05 (d, 4 H), 5.01 (dd, 4 H) ), 4.89 (dt, 4 H), and 4.19 - 4.22 ppm (m, 4 H) ; 13 C NMR (D2O, 100 MHz) δ 169.1, 143.1, 138.0, 137.3, 133.4, 128.2, 127.3, 125.0, 121.8, 99.3, 76 , 0.75,7, 73.4, 72.5, 70.8 and 67.7 ppm. Anal. Calc. for C54H52N2074Si6Na16 16 H2 O: C, 21.01; H, 2.75; N, 0.91; S, 16.65. Found: C, 20.93; H, 2.78; N, 0.77; S, 16.86. Capillary electrophoresis showed a purity greater than 91%. EXAMPLE 18

Step 1

To a solution of 3,5-bis (hepta-O-acetyl-β-D-maltosyloxymethyl) phenylamine hydrochloride A solution of 3,5-bis (hepta-O-acetyl-β-D-maltosyloxymethyl) phenylamine (1.07 g, 0.769 mmol) and triethylamine (101 æL, 0.769 mmol) in THF (20 mL) was added 4,4-biphenyldicarboxylic acid dichloride (107 mg, 0.385 mmol). H, the reaction mixture was quenched with MeOH, diluted with CH 2 Cl 2 and washed with H 2 O. The reaction mixture was dried (MgSO 4) and concentrated to give 1.10 g (96% yield) of the title compound as a colorless solid, mp NMR: (CDCl 3, 400 MHz) δ 8.66 (broad s, 2 H), 8.03 (d, 4 H), 7.72 (d, 4 H), 7.67 (s, 4 H), 6.93 (s, 2 H), 5.39 (d, 4 H), 5.32 (t, 4 H), 5.20 (t, 4 H), 5.02 (m, 12 H), 4.57 - 4.66 (m, 12 H), 4.20 - 4.25 (m, 8 H), 3 , 93-4.04 (m, 12 H), 3.6-3.7 (m, 4 H), 2.09 (s, 12 H), 2.06 (s, 12 H), 1.99 (s, 12 H), 1.98 (s, 12 H), 1.97 (s, 12 -43 - Η), 1.96 (s, 12 Η) and 1.95 ppm (s, 12 H) IR (KBr) 1745 cm -1. Anal. Calc. Calc'd for C 13 H 16 4 N 2 O 74: C, 53.89; H, 5.53; N, 0.94. Found: C, 53.49; H, 5.55; N, 0.94.

Step 2

Bis (3,5-bis (B-D-maltosyloxymethyl) phenylamide) biphenyl-dicarboxylic acid

A solution of biphenyl-4,4'-dicarboxylic acid bis {[3,5-bis (hepta-O-acetyl-D-maltosyloxymethyl) phenyl] amide} and 9.77 mL (9.37 mmol) , 77 mmol) of 1N NaOH in MeOH (15 mL) was stirred at 50 ° C for 4 hours. The reaction mixture was cooled to room temperature, quenched with 1N HCl (9.12 mL), concentrated and purified by chromatography on Saphadex G-10 (elution with H2 O). Removal of H2 O in vacuo gave the title compound, 494 mg (75% yield), mp 200 ° C (dec.): 1 H-NMR (D2 O, 400 MHz) δ 7.54 (s, 4 H ), 7.33 (s, 4 H), 7.23-7.27 (broad s, 4 H), 7.15 (s, 2 H), 5.32 (s, 4 H), 4.51 (D, 4 H), 3.50-3.90 (m, 36 H), and 3.30-3.45 ppm (m, 12 H) ; , Δ 166.3, 141.7, 137.8, 137.3, 132.0, 127.7, 126.5, 124.1, 120.1, 101.3, 99.9, 77.4, 75.9, 74.4, 72.8, 72.6, 71.6, 70.6, 69.1, 60.6 and 60.3 ppm. Anal. Calc. Calc'd for C 78 H 108 N 2 O 46 8H 2 O: C, 48.85; H, 6.31; N, 1.46. Found: C, 48.48; H, 6.28; N, 1.58.

Step 3

Biphenyl-4,4'-dicarboxylic acid bisf (3,5-bis (hepta-β-sulfato-β-D-maltosyloxymethyl) phenyl) ammonium salt

A solution of 317 mg (0.174 mmol) of biphenyl-4,4'-dicarboxylic acid bis {[3,5-bis (pD-4-maltosyloxymethyl) phenyl] amide and 3.58 g (25.7 mmol ) of sulfur trioxide complex and trimethylamine in DMF (15 mL) was stirred at 70 ° C for 3 days. The reaction mixture was cooled to room temperature, quenched with distilled H2 O and concentrated. The residue was purified by passage through a Sephadex G-10 column (elution with H 2 O). Exchange of cations was achieved by passing an aqueous solution of the product through a Dowex 50 x 8 column of strongly acidic resin (Na-form). Removal of the solvent and azeotroping with toluene gave 609 mg (75%) of an off-white solid, mp 178øC (dec.): 1 H-NMR (D2 O, 400 MHz) δ 8.06 (d, 4 H ), 7.94 (d, 4 H), 7.69 (s, 4 H), 7.36 (s, 2 H), 5.58 (d, 4 H), 5.06 (d, 4 H) ), 5.02 (d, 4 H), 4.7-4.9 (m, 12 H), 4.58-4.61 (m, 8 H), 4.49 (dd, 4 H), 4.12-4.46ppm (m, 28H); 13 C NMR (D2O, 100 MHz) δ 169.1, 143.1, 138.0, 137.3, 133.4, 128.2, 127.4, 124.9, 121.7, 99.3, 94 , 1.77,1.76.0, 74.8, 73.2, 73.1, 72.3, 71.8, 70.5, 69.9, 67.5 and 66.0 ppm; mass spectrum (&quot; electrospray &quot;) (m-zNa) / z 1143.7 (m-4 Na) 4 ', 910.4 (m- 5 Na 5') 754.8 (m- 6 Na 6 ') 643, 7 (m-7 Na7 '), 560.4 (m-8 Na) 8'. Anal.

Calc. Calc'd for C 78 H 12 N 2 O 3 • S 28 Na 2 • 28 H 2 O: C, 18.12; H, 2.65; N, 0.54; S, 17.36. Found: C, 18.33; H, 2.73; N, 0.46; S, 17.72. EXAMPLE 19

Step 1: (R) -N-N-Ureidolbis-N-FS-theta-O-acetyl-β-D-cellobiosyloxymethyl)

chlorophenyl) benzamide A solution of 1.10 g (1.42 mmol) of 5- (hepta-O-acetyl-PD-cellobiosyloxymethyl) -2-chlorophenylamine in THF (20 mL) containing 198 μL of triethylamine (1.42 mmol) was added 3-nitrobenzoyl chloride (316 mg, 1.70 -45-

mmol). After 3 h, the reaction mixture was quenched with MeOH and diluted with CH 2 Cl 2. The reaction mixture was washed with H2 O, dried (MgSO4), filtered and concentrated to 1.31 g (1.42 mmol) of crude product which was used directly in the next reaction. The crude material was dissolved in EtOAc and treated with 2.32 g (10.28 mmol) of SnCl2 · H2 O. After stirring at reflux for 5 h, the reaction mixture was cooled to room temperature and quenched with sat. (300 mL). The reaction mixture was diluted with EtOAc and filtered. The organic phase was separated and the aqueous phase was extracted with CH 2 Cl 2. The organic phases were combined, dried (MgSO4) and concentrated to give 1.18 g (93% yield) of crude product which was directly used in the next reaction. To the THF product (20 mL) containing 108 æl (7.98 mmol) of pyridine was added 66 mg (0.22 mmol) of triphosgene. The mixture was stirred at room temperature overnight. The reaction mixture was quenched with H2 O and stirred for another 30 min. The reaction mixture was filtered. The solid was collected and dissolved in CH 2 Cl 2, dried (MgSO 4), concentrated and then triturated with Et 2 O to give 995 mg (82% yield) of the title compound, mp 173-178 ° C: 1 H-NMR (CDCl3, 400 1 H NMR (CDCl 3) δ 8.47 (s, 2 H), 8.41 (d, 2 H), 7.45 (d, 2 H), 7.38 (t, 2 H), 7.33 (d, 2 H), 6.96 (dd, 2 H), 5.10-5.16 (m, 4 H), 5.03 (t, 2 H), 4.90-4.96 (m, 4 H) , 4.79 (d, 2H), 4.59 (t, 2H), 4.51-4.56 (m, 4H), 4.30 (dd, 1H), 4.14 (dd, , 3.99 (dd, 2 H), 3.92 (t, 2 H), 3.65-3.70 (dq, 2 H), 2.09 (s, 6 H), 2 (S, 6 H), 1.985 (s, 6 H), 1.96 (s, 6 H), 1.96 (s, 6 H); 13 C NMR (CDCl 3, 100 MHz) δ 170.6, 170.49, 170.46, 169.83, 169.17, 169.14, 165.22, 152.47, 139.75, 136.78, 134 , 85, 134.43, 129.68, 129.13, 124.35, 123.08, 122.95, 121.18, 120.57, 117.99, 100.70, 98.92, 73.22 , 72.64, 71.89, 71.74, 71.55, 71.49, 69.97, 67.90, 62.04, 61.53, 20.86, 20.72, 20.64, 20 , 58, 20.53 and 20.45 ppm; mass spectrum ((+) - FAB) m / z 1837.5 (M + Na). Anal. Calc. Calc'd for C 81 H 92 N 4 O 3 · Cl 2 • H 2 O: C, 53.03; H, 5.16; N, 3.05. Found: C, 52.74; H, 5.09; N, 3.15.

Step 2: 3,3 '- (1α-Ureidolbis (N- [5- (β-D-cellobiosyloxymethyl) -2-chlorophenyl] benzamide

A solution of 908 mg (0.50 mmol) of 3,3 '- (N, N'-Ureido) bis {N- [5- (hepta-O-acetyl-β-cellobiosyloxymethyl) -2-chlorophenyl] benzamide} in MeOH (20 mL) containing 7.5 mL (7.5 mmol) 1 N NaOH was stirred at 50 ° C under N 2 for 4 h. The reaction mixture was cooled to room temperature and quenched with 1N HCl (7.0 mL, 7.0 mmol). The reaction mixture was stirred for 15 min. and filtered. The solid was collected and azeotroped dry with toluene to give 566 mg (92% yield) of the title compound as an off-white solid, mp 183 ° C: 1 H-NMR (DMSO-d 6, 400 MHz) δ 10, (S, 1 H), 2 H), 7.75 (d, 1 H), 7.60 (d, 2 H), 7 (S, 2 H), 7.50-7.55 (q, 4 H), 7.44 (d, 2 H), 7.33 (dd, 2 H), 5.30 (s, 2 H), 5.25 (d, 2H), 4.86 (d, 2H), 4.70 (s, 2H), 4.33 (d, 2H), 4.25 H), 3.75-3.80 (dd, 2 H), 3.60-3.71 (m, 4 H) and 3.0-3.2 ppm (m, 8 H); 13 C-NMR (DMSO-d 6, 100 MHz) δ 165.54, 152.70, 140.05, 137.74, 134.90, 134.78, 129.27, 128.95, 128.48, 127.36 , 126.53, 121.58, 120.91, 117.80, 103.21, 101.99, 80.49, 76.79, 76.46, 75.01, 73.29, 73.19, 70 , 04, 68.82, 61.02 and 60.43 ppm; mass spectrum ((+) - FAB) m / z 1249.2 (M + Na). Anal. Calc. Calc'd for C 24 H 24 N 4 O 3 · 4HCl: C, 48.97; H, 5.58; N, 4.31. Found: C, 49.25; H, 5.58; N, 4.28.

Step 3

3.3 t- (N, N -ureidolbis ((N - [2-chloro-5- (hepta-4-sulfatoethyl) -3-D-cellobiosyloxymethyl) phenyl] benzamid

A solution of 3,3 '- (N, N'-ureido) bis {N- [5- (β-cellobiosyloxymethyl) -2-chlorophenyl] benzamide} (475 mg, 0.387 mmol) in DMF (25 mL) containing 3 , (27.08 mmol) of sulfur trioxide and trimethylamine complex was stirred at 70 ° C for 3 days. The reaction mixture was cooled to room temperature, quenched with distilled H2 O and concentrated. The residue was purified by passage through a Sephadex G-10 column (elution with H 2 O). Exchange of cations was accomplished by passing an aqueous solution of the product through a Dowex 50 x 8 column of strongly acidic resin (Na-form). Removal of the solvent and azeotroping with toluene gave 926 mg (0.349 mmol) of the title compound as an off-white solid, mp 168øC (dec.): 13 C NMR (D2 O, 100 MHz) δ 169.46, 29, 138.29, 136.85, 133.89, 133.06, 129.90, 129.63, 129.56, 128.43, 127.62, 124.74, 122.79, 119.59, 99.91, 99.14, 77.61, 77.32, 77.03, 74.28, 73.63, 73.34, 70.0, 67.60 and 66.60 ppm. Anal. Calc. Calc'd for C 55 H 58 N 4 O 6 S 4 S 4 N 4 O 4 Na 2 SO 4 184H 2 O: C, 19.08; H, 2.60; N, 1.68; S, 16.34. Found: C, 19.01; H, 2.21; N, 1.74; S, 16.82. Capillary zone electrophoresis showed purity &gt; 90%. EXAMPLE 20

Step 1

To a solution of 5- (hepta-O-acetyl-PD-cellobiosyloxymethyl) -2-chlorophenylamine (995 mg, 1.28 mmol), and (R) -hexahydro- triethylamine (148 æL, 1.28 mmol) in THF (25 mL) was added 4,4-biphenyldicarboxylic acid dichloride (179 mg, 0.642 mmol). After stirring at room temperature for 2 h, the reaction mixture was quenched with MeOH, diluted with CH2 Cl2 and washed with H2 O. The reaction mixture was dried (MgSO4) and concentrated to an oil which was triturated with ether to give 1.09 g of crude product (99% yield). dissolved in MeOH (20 mL) and treated with 9.5 mL of 1 N NaOH. After stirring at 50 ° C for 4 h, the reaction mixture was cooled to room temperature and treated with 8.9 mL (8.9 mmol ) of 1N HCl. Removal of the solid afforded 670 mg (91% yield) of the title compound, mp> 200 ° C: 1 H-NMR (DMSO-d6, 400 MHz) δ 10.21 (s, 2 H), 8.13 (d, 4 H), 7.95 (d, 2H), 7.35 (d, 2H), 7.35 (dd, 2H), 5.29 (d, 2H), 5.25 -5.26 (broad, 2 H), 4.87 (d, 2 H), 4.71 (s; 2 H), 3.78 (dd, 2 H), 3.15 (broad d, 2 H), and 3.05 ppm (d, 2 H) t, 2 H); 13 C NMR (DMSO-d 6, 100 MHz) δ 165.0, 128.5, 127.0, 103.2, 102.0, 80.5, 76.8, 76.4, 75.0, 73.3 , 73.2, 70.0, 68.8, 61.0 and 60.4 ppm; mass spectrum (-FAB) m / z 1166.9, 1004.9 and 842.9. Anal. Calc. Calc'd for C 52 H 62 N 2 O 24 Cl 2 5H 2 O: C, 49.57; H, 5.76; N, 2.22. Found: C, 49.81; H, 5.36; N, 2.14.

Step 2

Bis (β-chloro-5- (hepta-β-sulfato-β-d-cellobiosyloxymethylphenylamide) bis (benzyl) dicarboxylic acid bis

A solution of 531 mg (0.454 mmol) of biphenyl-4,4'-dicarboxylic acid bis {[2-chloro-5- (PD-cellobiosyloxymethyl) phenyl] amide} and 4.42 g (31.8 mmol) of sulfur trioxide complex and trimethylamine in DMF (25 mL) was stirred at 70 ° C for 2 days. The reaction mixture was concentrated and purified by chromatography on a Sephadex G-10 column eluting with H2O. Cation exchange was performed using a Dowex 50 x 8 column of strongly acidic resin (Na-form) eluting with H2O. Removal of the water in vacuo and azeotroping with toluene gave 705 mg (60%) of the title compound as a white solid, mp 165øC (dec.): 1 H-NMR (DMSO-d 6, 400 MHz) δ 8 (D, 2H), 7.65 (d, 2H), 7.52 (dd, 2H), 7.96 (d, 2H) , 99 (d, 2H), 4.97 (d, 2H)

Η), 4.58 (d, 2 Η) and 4.00-4.06 ppm (m, 4 H); , Δ 169.8, 143.5, 137.0, 133.2, 132.6, 130.04, 130.01, 128.7, 128.3, 128.0, 127.5, 99.9, 99.2, 77.7, 77.4, 77.1, 74.3, 73.6, 73.4, 73.2, 67.7, and 66.6 ppm; mass spectrum (&quot; electrospray &quot;) (m-zNa) / z 496.7 (m-5 Na) 5 ', 626.6 (m- 4 Na) 4' and 843.2 (m- '. Anal. Calc. for C52H48Cl2N2Na14066Si416H2O: C, 21.63; H, 2.79; N, 0.97; S, 15.55. Found: C, 21.24; H, 2.31; N, 0.97; S, 12.25. EXAMPLE 21

Step 1

To a solution of 861 mg (1.11 mmol) of 2- (hepta-O-acetyl-PD-cellobiosyloxymethyl) -4-chlorophenylamine and 861 mg (1.11 mmol) of triethylamine (155 æL, 1.11 mmol) in THF (20 mL) was added 4,4-biphenyldicarboxylic acid dichloride (155 mg, 0.555 mmol). After stirring at room temperature for 2 h, the reaction mixture was quenched with MeOH, diluted with CH 2 Cl 2 and washed with H 2 O. The organic phase was dried (MgSO4), filtered and concentrated to give 975 mg of crude product. The material was dissolved in MeOH (15 mL) and treated with 8.3 mL (8.3 mmol) of 1 N NaOH. After stirring for 4 h. at room temperature, the reaction mixture was quenched with 7.8 mL of 1N HCl and the solid collected to provide 590 mg (91% yield) of the title compound as an off-white solid, mp> 200 ° C: 1 H-NMR (DMSO-d 6, 400 MHz) δ 9.95 (s, 2 H), 8.11 (d, 4 H), 7.93 (d, 4 H), 7.67 (d, 2 H), 4.62 (d, 2 H), 4.42 (dd, 2 H), 5.45 (d, 2 H), 4.91 (d, 2 H) 2 H) and 4.25 ppm (d, 2 H); 13 C partial-DMSO (DMSO-d 6, 100 MHz) δ 164.9, 142.2, 134.6, 134.5, 133.4, 128.5, 127.0, 103.2, 101.7 , 80.4, 80.2, 76.8, 76.4, 75.0, 74.9, 73.3, 70.0, 66.3, 61.0 and 60.3 ppm; IR (KBr) 1650 cm -1; mass spectrum (-FAB) m / z 1167.3, 843.3 and 704.2. Anal. Calc. Calc'd for C 52 H 62 Cl 2 N 2 O 244 H 2 O: C, 50.29; H, 5.68; N, 2.26. Found: C, 50.37; H, 5.38; N, 2.32.

Step 2

Tetradecodicarboxylic acid bis (4-chloro-2- (hepta-β-sulfato-B-D-cellobiosyloxymethyl)

A solution of 447 mg (0.382 mmol) of biphenyl-4,4'-dicarboxylic acid bis {[2- (PD-cellobio-siloxymethyl) -4-chlorophenyl] amide and 3.85 g (27.7 mmol) of sulfur trioxide complex and trimethylamine in DMF (25 mL) was stirred at 70 ° C for 6 days. The reaction mixture was quenched at ambient temperature with H2 O, concentrated and passed through a Sephadex G-10 column (elution with H2 O). Cation exchange was effected by passing an aqueous solution of the compound through a Dowex 50 x 8 column of strongly acidic resin (Na-form) eluting with H2O. Removal of the solvent and azeotroping with toluene gave 666 mg (67% yield) of the title compound as an off-white solid, m.p. 172 ° C which was ca. 70% pure as determined by capillary electrophoresis: â € ƒâ € ƒâ € ƒ1 H-NMR (D2 O, 400 MHz) , 7.46-7.53 (broad, 4 H), 4.95 (d, 2H) and 4.90 ppm (d, 2H); 1 H NMR (D 2 O, 100 MHz) δ 169.9, 143.4, 134.5, 133.4, 132.4, 129.7, 129.0, 128.7, 128.3, 127.6, 9.98.8, 77.6, 77.4, 77.0, 74.3, 73.5, 73.4, 73.0, 67.6, 66.9 and 66.3 ppm; Anal. Calc. Calc'd for C 52 H 48 Cl 2 N 2 Na 40 66 Si4'14 H 2 O: C, 21.92; H, 2.69; N, 0.98; S, 15.75. Found: C, 21.98; H, 2.63; N, 1.08; S, 16.05. EXAMPLE 22

To a solution of 840 mg (0.604 mmol) 3,5-bis (hepta-O-acetyl-β-D-cellobiosyloxymethyl) phenylamine and 86.4 μΐ. (0.31 mmol) triethylamine in THF (20 mL) was added succinyl chloride (34.2 μL, 0.31 mmol). After stirring at room temperature for 1 h, it was observed that the reaction had not been completed. Another 10 æl (0.09 mmol) of succinyl chloride was added. The reaction was stirred for another 15 min, quenched with MeOH, diluted with CH2 Cl2, washed with H2 O, dried (MgSO4), filtered and concentrated. Trituration with Et 2 0 gave a green solid which was semi-purified by flash chromatography &quot; (EtOAc) to give a yellow solid (633 mg, 75% yield). The compound was dissolved in MeOH and treated with 6.6 mL of 1N NaOH (6.6 mmol). After stirring at 50 ° C for 4 h, the reaction mixture was cooled to room temperature and strongly acidic Amberlite resin was added until a neutral pH was obtained. After stirring for 5 min, the reaction mixture was filtered and concentrated to a yellow solid. Purification by reverse phase chromatography (silica gel RP-60) eluting with MeOH: H 2 O (3: 7) and then chromatography with Sephadex G-10 (eluting with H 2 O) gave 278 mg (94%) of the title compound as a white solid. yellow solid: 1 H-NMR (δ, 400 MHz) δ 7.48 (s, 4 H), 7.31 (s, 2 H), 4.91 (d, 4 H), 4.71 (D, 4 H), 3.74 (dd, 4 H), 3.43 (d, 4 H), and 2.81 ppm ( s, 4H); 13 C NMR (D2O, 100 MHz) δ 173.3, 137.9, 137.3, 124.9, 120.9, 102.4, 101.0, 78.5, 75.8, 75.3, 74 , 6.74, 73.0, 72.7, 70.7, 69.3, 60.4, 59.9 and 31.4 ppm.

Step 2

N, N-bis-bis-3,5-bis (hepta-β-sulfato-β-D-cellobiosyloxymethylphenylsuccinamide

A solution of 278 mg (0.208 mmol) of N, N'-bis [3,5-bis (pD-cellobiosyloxymethyl) phenyl] succinamide and of sulfur trioxide and trimethylamine complex (4.23 g, 29.0 mmol) in DMF (20 mL) was stirred at 70 ° C for 3 days. The reaction mixture was quenched at room temperature with H2 O and then concentrated. Purification by chromatography on Sephadex G-10 (elution with H 2 O) and then exchange of cations using a Dowex 50 x 8 column of strongly acidic resin (Na-form) afforded 606 mg (64% yield) of the title compound as a white solid. brown solid, mp 168øC (dec.): 1 H-NMR partial (D 20, 400 MHz) δ 7.58 (s, 4 H), 7.36 (s, 2 H), 4.00-5.00 (m, 64 H) and 2.87 ppm (s, 4 H); 13 C NMR (D2O, 100 MHz) δ 173.3, 138.0, 137.1, 124.4, 99.9, 99.7, 77.6, 77.4, 77.0, 74.3, 73 , 4.72.9, 70.9, 67.6, 66.2 and 37.1 ppm; mass spectrum (&quot; electrospray &quot;) (m-zNa) / z 734.1 (m-6 Na) 6 ', 885.5 (m-5 Na) 5' and 1112.7 (m -4 Na) 4 '. Anal. Calc. Calc'd for C 68 H 76 N 2 O 13 () S 28 Na 2828 H2 O: C, 15.62; H, 2.93; N, 0.54; S, 17.18. Found: C, 15.24; H, 2.43; N, 0.75; S, 17.67. EXAMPLE 23

To a solution of 3,5-bis (hepta-O-acetyl-PD-cellobiosyloxymethyl) -phenylamine (823 mg, 0.592 mmol) and triethylamine (82 mg, 0.592 mmol) in dichloromethane 5 pL, 0.592 mmol) in THF (15 mL) was added terephthaloyl dichloride (60.1 mg, 0.296 mmol)

stirring at room temperature for 2 h, the reaction mixture was quenched with MeOH, diluted with CH 2 Cl 2 and washed with H 2 O. The organic phase was dried (MgSO4), filtered, concentrated and triturated with Et2 O to give 845 mg (98%) of crude product. This was dissolved in MeOH (20 mL) and treated with 8.7 mL (8.7 mmol) of 1 N NaOH. After stirring at 50 ° C for 3 h, the reaction mixture was cooled to room temperature, was added 8.1 mL (8.1 mmol) of 1 N HCl. The solid was collected and dried in vacuo to give 295 mg (59% yield) of the title compound as a white solid, mp 185øC (dec. ): 1 H-NMR (DMSO-d 6) δ 8.01 (s, 4 H), 7.64 (s, 4 H), 7.18 (s, 2 H) , 4.31 (d, 2H), 4.56 (d, 2H), 4.33 (d, 2H), 4.26 (d, 2H), and 3.77 ppm H); 13 C NMR. (DMSO-d 6, 100 MHz) δ 164.7, 138.7, 138.2, 137.3, 127.7, 122.9, 119.3, 103.2, 101.9, 80.5, 76 , 8, 76.4, 75.1, 75.0, 73.3, 73.2, 70.0, 69.9, 61.0 and 60.4 ppm; IR (KBr) 1650 cm -1. Anal. Calc. Calc'd for C 72 H 104 N 6 O 46: 2 H 2 O: C, 44.35; H, 6.62; N, 1.44. Found: C, 44.05; H, 6.30; N, 1.41.

Step 2

N-bis-3,5-bis (hepta-β-sulfato-β-D-cellobiosyloxymethyl) phenyl trimethylphthalamide octacosodium salt

A solution of 178 mg (0.103 mmol) of N, N'-bis [3,5-bis (PD-cellobiosyloxymethyl) phenyl] terephthalamide and of sulfur trioxide and trimethylamine complex (2.0 g, 14.4 mmol) in DMF (20 mL) was stirred at 70 ° C for 3 days. The reaction mixture was cooled, quenched with water and concentrated. The residue was purified by chromatography on Sephadex G-10 (elution with H2 O). Exchange of cations was effected using a Dowex 50 x 8 column of strongly acidic resin (Na-form) afforded 402 mg (85% yield) of the title compound as an off-white solid, mp 163øC (dec.): 1 H-NMR (D 2 O, 400 MHz) δ 8.09 (s, 4 H), 7.71 (s, 4 H), 7.43 (s, 2 H), 5.03 (d, 2 H) and 5.00 ppm -54- (d, 2H); 13 C NMR (DMSO-d 6, 100 MHz) δ 168.7, 138.1, 137.2, 137.1, 127.8, 125.0, 121.7, 99.9, 99.7, 77.6 , 77.4, 77.0, 74.3, 73.4, 72.9, 70.9, 67.6 and 66.3 ppm; mass spectrum (&quot; electrospray &quot;) (m-zNa) / z 632.8 (m-7Na) 7 ', 742.1 (m -6Na) 6 &quot;, 895.2 (m- 5Na) 5' and 1124 , 7 (m-4) 4. Calc. for C72H76N2

Na280.3, 288 28 H2O: C, 16.97; H, 2.61; N, 0.55; S, 17.62. Found: C, 16.77; H, 2.41; N, 0.59; S, 17.42. EXAMPLE 24

Step 1

Bisir5- (B-D-cellobiosyloxymethyl) -2-methylpheninamide &gt; To 970 mg (1.28 mmol) of 5- (hepta-O-acetyl-PD-cellobiosyloxymethyl) -2-methylphenylamine in CH 2 Cl 2 (20 mL) containing 1.0 mL (12.4 g) mmol) of pyridine was added 225 mg (0.641 mmol) of biphenyl-4,4'-disulfonyl chloride. After stirring at room temperature for 90 min, the reaction mixture was quenched with sat. and extracted into EtOAc. The bright yellow organic phase was washed with pH 7 buffer, dried (MgSO4 and flash chromatographed Et0Ac / Et2O / CH2Cl2, 1: 1: 1) to give 504 mg (58% yield) of a powder of Orange. This material (a total of 675 mg (0.378 mmol) of two separate procedures) was dissolved in MeOH (10 mL and THF (10 mL) and treated with 6 mL of 1 N NaOH. After stirring at room temperature for 12 h, the reaction mixture was quenched with 6 mL of 1N HCl and concentrated to ca 6-8 mL The resulting precipitate was collected and washed with 15 mL of H2 O and then Et2 O. The azeotropic drying with toluene gave 335 mg 1 H-NMR (DMSO, 400 MHz) δ 9.69 (s, 2 H), 7.91 (d, 4 H) , 7.75 (d, 4 H), 7.16 (d, 2-55-

Η), 7.11 (s, 2 Η), 7.10 (d, 2 Η), 4.71 (d, 2 Η), 4.46 (d, 2 Η), 4.24-4.28 , 3.77 (d, 2 Η), 3.70 (d, 2 Η), 3.58-3.64 (m, 2 Η), 2.97-3.43 (m, 18 Η) and 1 , 91 ppm (s, 6 H); 13 C NMR (DMSO, 100 MHz) δ 142.2, 136.1, 134.4, 132.9, 130.5, 127.8, 127.2, 126.0, 103.2, 101.6, 80 , 76.7, 76.4, 75.1, 74.9, 73.2, 73.1, 70.0, 69.1, 61.0, 60.4 and 17.2 ppm; mass spectrum ((-) - FAB), m / z 1199.3, 750.2, 514.2. Anal. Calc. Calc'd for C 52 H 68 N 2 O 5 S 2 H 2 O: C, 51.23; H, 5.79; N, 2.30. Found: C, 51.04; H, 5.64; N, 2.29.

Step 2

Bis ({5- [p-D-cellobiosyloxymethyl) -2-methylphenyl] -2-methyl-5- (hepta-β-sulfato-BD-cellobiosyloxymethylphenylamide) bis ] amide hydrochloride in DMF (10 mL) was added 1.8 g (12.9 mmol) of trimethylamine sulfur trioxide complex After stirring for 3 days at 70 ° C, the reaction mixture was cooled to room temperature, quenched with 10 mL of water and stirred for 20 min. This was concentrated in vacuo and purified by gel filtration (Sephadex G-10) to give 419 mg of light brown flakes. was dissolved in a minimum amount of water and passed through an ion exchange column (Dowex 50 x 8 strongly acidic (Na-form)) to give 350 mg of light brown flakes after azeotroping with toluene: 1 H-NMR (D2 O, 400 1H NMR (CDCl3) δ 7.89 (d, 4 H), 7.81 (d, 4 H), 7.39 (d, 2 H), 7.32 (s, 2 H), 7.25 H), 4.94 (d, 2H), 4.65-4.87 (m, 8 H), 4.57 (dd, 2 H), 4.18-4.51 (m, 16 H), 3.98-4.05 (m, 4 H) and 1.90 ppm (s, 6 H); 13 C NMR (D2O, 100 MHz) δ 143.8, 138.0, 135.4, 134.9, 131.1, 128.1, 128.0, 127.8, 127.3, 99.9.98 , 7.77.6, 77.3, 77.2, 76.9, 74.3, 73.7, 73.3, 72.93, 70.3, 67.6, 66.6 and 16.4 ppm. Anal. Calc. Calc'd for C 52 H 54 N 2 Nal 4068 Si 6 18 H 2 O: C, 21.14; H, 3.05; N, 0.95. Found: C, 21.17; H, 2.71; N, 0.96. -5-

Example 25

To a solution of 879 mg (1.88 mmol) of 5- (tetra-O-acetyl-PD-glucopyranosyloxymethyl) -2-methylphenylamine and 262 Âμl of N, (1.88 mmol) of triethylamine in THF (10 mL) was added succinyl chloride (104 μL, 0.94 mmol). After stirring at room temperature for 3 h, the reaction mixture was quenched with MeOH (20 mL), diluted with CH 2 Cl 2, washed with H 2 O, dried (MgSO 4) and concentrated. Crystallization from Et2 O gave 741 mg (78%) of a green solid which was directly used in the next reaction. To a solution of 360 mg (0.354 mmol) of the crude product in MeOH (7.5 mL) was added 1N NaOH (3.54 mL, 3.54 mmol). After stirring at 50 ° C for 90 min, the reaction mixture was quenched at ambient temperature with 1N HCl (2.85 mL, 2.50 mmol). After stirring for 1 h. at room temperature the solid was collected to give 229 mg of the title compound: 1 H-NMR (DMSO-d 6, 300 MHz) δ 9.41 (s, 2 H), 7.36 (s, 2 H) (D, 2 H), 4.78 (d, 2 H), 4.50 (d, 2 H), 4.22 (d, 2 H) , 3.69 (d, 2H), 2.67 (s, 4 H) and 2.18 ppm (s, 6 H).

Step 2

N, N, -bisr2-methyl-5- (2 &lt; 3 &gt; 4,6-tetra-Q-sulfato-β-D-glucopyranosyloxymethylphenylsuccinamide, octassodic salt

A solution of 229 mg (0.337 mmol) of N, N'-bis [2-methyl-5- (PD-glucopyranosyloxymethyl) phenyl] succinamide and sulfur trioxide trimetylamine complex (2.11 g, 15.2 mmol ) in DMF (30 mL) was stirred at 50 ° C overnight. The reaction mixture was quenched with H2 O (10 mL) and concentrated to an oily solid. The solid was removed and washed with a very small amount of H2 0. The combined filtrates were concentrated and partially purified by repeated chromatography on Sephadex G-10 (elution with H2 O) and then by dialysis using a 500 MW cut bag. A small amount of trimethylammonium sulfate was further found by NMR. The cations were exchanged with a Dowex 50 x 8 column of strongly acidic resin (Na-form) (elution with H2 O) to provide 249 mg (49% yield) of the title compound as an off-white solid, m.p. 200 ° C: Partial1 H-NMR (D2 O, 400 MHz) δ 7.39 (d, 2H), 7.37 (d, 2H) and 4.55 ppm (t, 2H); 13 C NMR (D2O, 100 MHz) δ 173.9, 134.82, 134.80, 134.3, 130.8, 127.6, 98.8, 75.9, 75.5, 73.4, 72 , 4.70, 67.8, 31.0 and 16.7 ppm; mass spectrum ((-) FAB) m / z 1472.6 (M-Na) +, 1370.7 (m-Na-NaS03 + H) +, 765.9, and 564.8. Anal. Calc. for C32H36N2Na8038S8'2NaS046 H2O: C, 20.34; H, 2.54; N, 1.48. Found: C, 20.19; H, 2.22; N, 1.39. EXAMPLE 26

To a stirred mixture of 5- (Hepta-O-acetyl-β-maltosyloxymethyl) -2-methylphenylamide (0.8 g) in dichloromethane 1.90 g, 2.52 mmol) and pyridine (0.22 g, 2.77 mmol) in CH 2 Cl 2 (10 mL) was added 3-nitrobenzoyl chloride (0.51 g, 2.77 mmol). The mixture was diluted with EtOAc, washed with sat. NaHCO3, brine, dried (MgSO4), and concentrated. Flash chromatography (1: 1 EtOAc / hexane) afforded 2,2,4- (DMSO-d6, 400 MHz) δ 10.28 (s, 1 H), 8.80 (s, 1 H) , 8.45 (dd, 1 H), 8.42 (d, 1 H), 7.84 (m, 1 H), 7.28 (d, 1 H), 7.25 (s, 1 H) (D, 1 H), 5.30 (d, 1 H), 5.05 (t, 1 H), 4.85 (d, 1 H) , 4.80 (m, 1 H), 4.75 (m, 2 H), 4.56 (d, 1 H), 4.40 (dd, 1 H), 4.19 (m, 2 H) , 4.0 (m, 4 H), 2.20 (s, 3 H), and 2.00 ppm (m, 21 H) Calc'd for C41 H48 N2 O21: C, 54.42; H, 5.35 N, 3.10 Found: C, 54.30, H, 5.27; N, 3.10.

To a stirred solution of N- [5- (hepta-O-acetyl-PD-malto-siloxy) -2-methylphenyl] -3-nitrobenzamide nitrobenzamide (2.00 g, 2.21 mmol) in MeOH (20 mL) was added 25% by weight of NaOMe in MeOH (5.10 mL, 22.10 mmol). After 3 h, 10 g of Amberlite IR-120 (H +) ion exchange resin The mixture was filtered and the filtrate concentrated to give 1.30 g (96%) of the title compound as a white solid, mp 166-168 ° C: 1 H NMR (DMSO-d 6, 400 MHz) δ 10.37 (s, 1 H), 8.80 (t, 1 H), 8.43 (m, 2 H), 7.85 (m, 1 H) (D, 1 H), 4.85 (d, 1 H), 4.57 (d, 1 H), 7.35 (s, 1 H) , 3.30 (m, 2 H), 3.30 (m, 2 H), 3.40 (m, 6 H), 3.10 (m, 2 H) and 2.21 ppm (s, 3H), mass spectrum m / e 609 (MH).

Step 3

N-R5- (Hepta-O-suphate-B-D-maltosyloxy) -2-methylphenyl] -3-nitrobenzamide heptaddicate salt

A mixture of N- [5- (PD-Maltosyloxymethyl) -2-methylphenyl] -3-nitrobenzamide (1.27 g, 2.08 mmol) and trimethylamine sulfur trioxide complex (10.13 g , 73.79 mmol) in DMF (10 mL) was heated at 70-75 ° C for 4 days. The mixture was cooled, MeOH was added and the mixture was filtered. The concentrated filtrate, taken up in MeOH and passed through a Sephadex LH-20-100 column (1: 1 MeOH / CHCl 3: 0.1% Et 3 N). The fractions were concentrated and passed through a Sephadex G-10 gel filtration column (H 2 O) to give a brown oil. Further purification by reverse phase chromatography (C-18; H 2 O), flash chromatography &quot; (5: 0.1: 0.5 CH 3 CN / H 2 O / Et 3 N) and ion exchange in Sephadex SP C-25 (Na +) provided 1.10 g (40%) of the title compound as a white solid, mp 168-170 Δ 8.82 (t, 1 H), 8.52 (m, 1 H), 8.38 (m, 1 H), 7.86 (m, 1 H), 7.45 (s, 2 H), 5.57 (d, 1 H), 4.99 (d, 1 H), 4.80 (m, 2 H), 4.62 (t, 1 H), 4.55 (s, 1H), 4.46 (m, 1 H), 4.30 (m, 4 H), 4.18 (m, 1 H), 4.05 (m, 1 H) and 2.31 ppm (s, 3H); mass spectrum m / z 1323. Anal. Calc. Calc'd for C 27 H 27 N 2 Na 7 O 7 OS 3: C, 23.52; H, 2.41; N, 2.03. Found: C, 22.70; H, 2.69; N, 1.72.

Step 4

3-N- [5- (hepta-O-sulfato-β-D-maltosyloxy) -2-methylphenyl] phenylamide

A solution of N- [5- (hepta-O-sulfato-PD-maltosyloxy) -2-methylphenyl] -3-nitrobenzamide (0.60 g, 0.45 mmol) in MeOH, H2 O and pyridine was hydrogenated at atmospheric pressure over 10% Pd / C for 6 h. The catalyst was removed by filtration and the filtrate was concentrated and passed through a Sephadex SP-C25 (Na +) ion exchange column to give 0.40 g (68%) of the title compound as a white solid, mp 232-234 1 H NMR (D 2 O, 300 MHz) δ 7.20 (m, 6 H), 6.93 (m, 1 H), 5.42 (d, 1 H) (M, 2 H), 4.40 (dd, 2 H), 4.30 (t, 1 H), 4.75 (m, 2 H) 15 (m, 2 H), 4.05 (m, 3 H), 3.95 (m, 2 H) and 2.15 ppm (s, 3 H). -60-

Step 5

To a stirred mixture of 3- [N- [5-methyl-5- (hepta-O-sulfato-BD-maltosyloxymethyl) phenyl] benzamide To a stirred mixture of 3- [N- [5- (440 mg, 0.34 mmol) and pyridine (5 mL) was added triphosgene (20 mg, 0.057 mmol) .Stirring was continued at room temperature The mixture was concentrated, through a Sephadex G-10 gel filtration column, and a Sephadex SP-C25 (Na +) ion exchange column. The fractions were concentrated and triturated with MeOH to give 350 (795) of the title compound as an off-white solid, mp 220øC (dec.): 1 H-NMR (D2 O, 400 MHz) δ 7.61 (d, 2H), 7.59 (s, (D, 2H), 4.95 (m, 8H), 7.29 (d, 2H), 5.54 (d, 2H) 4H), 4.80 (m, 2 H), 4.61 (t, 2 H), 4.52 (dd, 2 H), 4.44 (t, 4 H), 4.30 H), 4.20 (m, 2 H), 3.98 (t, 2 H) and 2.28 (s, 6 H); 13 C NMR (D 2 O, 100 MHz) δ 169.8, 135.1, 134.8, 134.4, 131.1, 130.0, 128.1, 126.9, 122.3, 121.7, 117.6, 94.7, 77.6, 76.6, 75.0, 73.7, 73.3, 1, 72.5, 69.8, 67.8, 6.0 and 16.8 ppm; mass spectrum m / z 1797. Anal. Calc. Anal. Calc'd for C 18 H 18 N 4 O 6 SOS 18H 2 O: C, 22.47; H, 3.15; N, 1.91. Found: C, 22.11; H, 2.81; N, 1.59. EXAMPLE 27

To a stirred solution of 3-amino [5- (hepta-O-acetyl-β-D-benzofuranyl) amide 2-methylphenyl] benzamide (1.0 g, 1.143 mmol) and triethylamine (0.12 g, 1.143 mmol) in CH2 Cl2 (5 mL) was added isophthaloyl dichloride (0.12 g, 0.572 mmol) . After 18 h, water was added and the phases were separated. The organic phase was washed with saturated aqueous NaHCO 3 solution and brine, dried (MgSO 4) and concentrated. Purification by flash chromatography &quot; afforded 0.57 g (53%) of product as a white solid, mp 178-180 ° C: δ 10.60 (s, 2 H), 9.90 (d, (s, 2H), 8.60 (s, 1 H), 8.37 (s, 2 H), 8.21 (dd, J = 1.5, 7.7 Hz, 2 H), 8.05 (d, J = 7.7 Hz, 2 H), 7.75 (m, 3 H), 7.52 (t, (M, 5H), 5.03 (m, 3 H), 4.80 (m, 4 H), 4.10 (d, J = 7.8 Hz, 2 H) (M, 4 H), 4.50 (d, J = 12.5 Hz, 2 H), 4.40 (d, J = 12.5 Hz, 2 H), 4.20 ), 3.95 (m, 8 H), 2.20 (s, 6 H) and 1.90-2.00 ppm (m, 42 H).

To a stirred solution of N, N'-bis {3- [2-methyl-5- (hepta-O-acetyl-PD-N-diisopropylcarboxamide) (0.572 g, 0.304 mmol) in MeOH (10 mL) was added 25 wt% NaOMe in MeOH (0.5 mL, 2.13 mmol). After 2 h, Amberlite IR-200 (H +) resin was added. The mixture was filtered and concentrated to afford 0.300 g (77%) of product as a white solid, mp 232-234 ° C: 1 H-NMR (DMSO-d6, 300 MHz) δ 10.65 (s, 2H) , 9.90 (s, 2 H), 8.60 (s, 1 H), 8.35 (s, 2 H), 8.21 (dd, J = 1.5, 7.7 Hz, 2 H (D, J = 7.7 Hz, 2 H), 7.70 (m, 2 H), 7.50 (t, J = 7.9 Hz, 2 H), 7.35 (d, (d, J = 12.5 Hz, 2 H), 4.80 (m, 5 H), 5.01 (d, J = 3.9 Hz, 2 H), 4.80 4.50 (d, J = 12.5 Hz, 2 H), 4.30 (d, J = 7.9 Hz, 2 H), 3.20-3.75 (m, 10 (m, 4 H), 2.20 (s, 6 H). Step 3

N, N-Bis [3- (2-methyl-5- (hepta-O-sulfato-3-D-maltosyloxymethylphenylcarbamoyl) isophthalamide

A mixture of N, N'-bis {3- [2-methyl-5- (PD-maltosyloxymethyl) phenylcarbamoyl] phenyl} isophthalamide (0.30 g, 0.232 mmol) and sulfur trioxide and pyridine complex ( 2.60 g, 16.267 mmol) in DMF (10 mL) was stirred at ambient temperature for 2 days. The mixture was concentrated, purified on a Sephadex LH-20-100 column (0.5% EtsN / DMF) and converted to a sodium salt with Sephadex SP-C-25 ion exchange resin to give 0.200 mg of product as a quasi-solid white solid, mp 222 ° C (dec.): δ 8.45 (s, 1 H), 8.35 (br. s, 2 H), 8.10 (broad s, (D, J = 7.9 Hz, 2 H), 7.85 (d, 7.9 Hz, 2 H), 7.75 (br. S, 2 H), 7.57 (s, 2 H), 7.35 (broad s, 5 H), 4.75 (m, 4 H), 4.45 (m, 4 H), 4.30 (m, 4 H), 4. (M, 4 H), 3.57 (m, 4 H), 3.28 (m, 8 H) and 2.20 ppm (s, 6 H); 13 C NMR (D2 O, 100 MHz) δ 160, 137, 134, 130, 128, 120, 102, 96, 75, 74, 72, 62, 56, 49, 18 ppm.

Lisbon, March 17, 2000

Official Agent of Industrial Property RUA VICTOR CORDON, 14 1200 LISBOA

Claims (24)

A compound of formula I wherein each R 1, R 2, R 3 and R 4 are independently H, SO 3 M or a sugar group of the structure: and each oligosaccharide group having the structure contains 1 to 3 sugar groups; M is lithium, sodium, potassium or ammonium; n is 1 or 2; X is hydrogen, halogen, lower alkyl of 1 to 6 carbon atoms or lower alkoxy of 1 to 6 carbon atoms; Y is carbonyl or sulfonyl; Z is alkyl of 1 to 12 carbon atoms, or X is X as defined above; or a pharmaceutically acceptable salt thereof. wherein each R 1, R 1, R 3 and R 4 are independently H, SQ 3 M or: A compound according to claim 1 of formula I- and each oligosaccharide group of structure contains 1 or 2 sugar groups; M is lithium, sodium, potassium or ammonium; n is 1 or 2; X is hydrogen, halogen, lower alkyl of 1 to 6 carbon atoms or lower alkoxy of 1 to 6 carbon atoms; Y is carbonyl or sulfonyl; Z is alkyl of 1 to 12 carbon atoms, or a pharmaceutically acceptable salt thereof. A compound according to claim 2 which is the dodecanedioic acid bis {[2-methyl-5- (hepta-O-sulfato-PD-cellobiosyloxymethyl) phenyl] amide} salt or a pharmaceutically acceptable salt thereof. pharmaceutically acceptable salt. A compound according to claim 2 which is the N, N'-bis [5- (hepta-O-sulfato-PD-cellobiosyloxymethyl) -2-methyl-phenyl] -terephthalamide tetradecodicarbonic salt or a pharmaceutically acceptable salt thereof . A compound according to claim 2 which is the tetradecassodic salt of biphenyl-4,4'-dicarboxylic acid bis {[5- (hepta-O-sulfato-PD-cellobiosyloxymethyl) -2-methylphenyl] or a pharmaceutically acceptable salt thereof. A compound according to claim 2 which is the N, N'-bis [5- (hepta-O-sulfato-β-D-cellobiosyloxymethyl) -2-methyl-phenyl] isophthalamide tetradecodicarbonic salt or a pharmaceutically acceptable salt thereof. A compound according to claim 2 which is the sodium octacosodic salt of decaniodic acid bis {[3,5-bis (hepta-O-sulfato-β-D-cellobiosyloxymethyl) phenyl] -amide} or a pharmaceutically acceptable salt thereof. A compound according to claim 2 which is the octacosodium salt of biphenyl-4,4'-dicarboxylic acid bis {[3,5-bis (hepta-O-sulfato-PD-cellobiosyloxymethyl) phenyl] amide} or a pharmaceutically acceptable salt thereof. pharmaceutically acceptable salt. A compound according to claim 2 which is the sodium octacosodium salt of biphenyl-4,4'-dicarboxylic acid bis {[3,5-bis (hepta-O-sulfato-PD-lactosyloxymethyl) phenyl] or a pharmaceutically acceptable salt thereof. A compound according to claim 2 which is the sodium octacosodium salt of biphenyl-4,4'-bis (β- (hepta-O-sulfato-PD-maltosyloxymethyl) phenyl] dicarboxylic acid or a pharmaceutically acceptable salt thereof. A compound according to claim 2 which is the hexadecodic salt of biphenyl-4,4'-dicarboxylic acid bis {[3,5-bis (tetra-O-sulfato-β-D-glucosyloxymethyl) phenyl] or a pharmaceutically acceptable salt thereof. A compound according to claim 2 which is the tetradecassodic salt of biphenyl-4,4'-dicarboxylic acid bis {[2-chloro-5- (hepta-O-sulfato-PD-cellobiosyloxymethyl) phenylamide] or a pharmaceutically acceptable salt thereof; A compound according to claim 2 which is the tetradecassodic salt of biphenyl-4,4'-biphenyl-4-carboxylic acid {[4-chloro-2- (hepta-O-sulfato-PD-cellobiosyloxymethyl) phenyl] dicarboxylic acid or a pharmaceutically acceptable salt thereof. A compound according to claim 2 which is the N, N'-bis [3,5-bis (hepta-O-sulfato-PD-cellobiosyloxymethyl) -phenyl] succinamide octacosodicarbonic salt or a pharmaceutically acceptable salt thereof . A compound according to claim 2 which is the sodium, potassium salt of N, N'-bis [3,5-bis (hepta-O-sulfato-PD-cellobiosyloxymethyl) -phenyl] terephthalamide or a pharmaceutically acceptable salt thereof . A compound according to claim 2 which is the tetradecassodic salt of biphenyl-4,4'-disulfonic acid bis {[2-methyl-5- (hepta-0-sulfato-β-d-cellobiosyloxymethyl) phenyl] amide} a pharmaceutically acceptable salt thereof. A compound according to claim 2 which is the octassodic salt of N, N'-bis [2-methyl-5- (2,3,4,6-tetra-O-sulfatoethyl) -D-glucopyranosyloxymethyl) phenyl] succinamide or a pharmaceutically acceptable salt thereof. A compound according to claim 2 which is the 3,3 '- [1,3-ureido] bis [N- [2-methyl-5- (hepta-O-sulfato-PD-maltosyloxymethyl) phenyl]} benzamide or a pharmaceutically acceptable salt thereof. A compound according to claim 2 which is the 3,3 '- (N, N'-ureido) -bis - ({N- [2-chloro-5- (hepta-O-sulfate-PD -benzamide) or a pharmaceutically acceptable salt thereof. A compound according to claim 2 which is the N, N'-bis {3- [2-methyl-5- (hepta-O-sulfato-PD-maltosyloxymethyl) phenylcarbamoyl] phenyl} isophthalamide tetradecodicarboxylic salt or a pharmaceutically acceptable salt thereof. A compound of formula I in which X each R 1, R 2, R 3 and R 4 are independently H, SO 3 M or a sugar group having the structure: and each oligosaccharide group having the structure contains 1 to 3 sugar groups; M is lithium, sodium, potassium or ammonium; n is 1 or 2; X is hydrogen, halogen, lower alkyl of 1 to 6 carbon atoms or lower alkoxy of 1 to 6 carbon atoms; Y is carbonyl or sulfonyl; Z is alkyl of 1 to 12 carbon atoms, H H N X -8- and X is as defined above; or a pharmaceutically acceptable salt thereof; for use in the treatment of a human suffering from a condition which is characterized by excessive proliferation of smooth muscle. A compound for a use as claimed in Claim 21 wherein the disease or condition which is characterized by excessive proliferation of smooth muscle is restenosis. A pharmaceutical composition comprising an effective amount of a compound of formula I I wherein each R 1, R 2, R 3 and R 4 are independently H, SO 3 M or a sugar group of the structure: and each oligosaccharide group having the structure -9- % * - ssszss * iám8 * Λ XL Μ \ 0R2 contains 1 to 3 sugar groups; M is lithium, sodium, potassium or ammonium; n is 1 or 2; X is hydrogen, halogen, lower alkyl of 1 to 6 carbon atoms or lower alkoxy of 1 to 6 carbon atoms; Y is carbonyl or sulfonyl; Z is alkyl of 1 to 12 carbon atoms, and X is as defined above; or a pharmaceutically acceptable salt thereof. Use of a compound of formula I wherein each R 1, R 2, R 3 and R 4 are independently H, SO 3 M or a sugar group of the structure: and each oligosaccharide group having the structure contains 1 to 3 sugar groups; M is lithium, sodium, potassium or ammonium; n is 1 or 2; X is hydrogen, halogen, lower alkyl of 1 to 6 carbon atoms or lower alkoxy of 1 to 6 carbon atoms; Y is carbonyl or sulfonyl; Z is alkyl of 1 to 12 carbon atoms, X Η H and X is as defined above; or a pharmaceutically acceptable salt thereof; in the manufacture of a medicament for treating a human suffering from a condition which is characterized by excessive proliferation of smooth muscle. Lisbon, March 17, 2000 JORGE CRUZ Official Property Agent Industnat street VICTOR CORDON, 14 1200 LISBOA